US2003220472A1PendingUtilityA1
Interleukin-3 (IL-3) multiple mutation polypeptides
Priority: Nov 24, 1992Filed: Feb 19, 2002Published: Nov 27, 2003
Est. expiryNov 24, 2012(expired)· nominal 20-yr term from priority
Inventors:S. Christopher BauerMark Allen AbramsSarah Ruth Braford-GoldbergMaire Helena CaparonAlan Michael EastonBarbara Kure KleinJohn MckearnPeter O. OlinsKumnan PaikJohn Warren Thomas
A61K 38/00Y10S930/141C07K 2319/00A61P 7/00C07K 2319/02C07K 14/5403A61P 43/00C12N 15/11C07K 14/54
61
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Claims
Abstract
The present invention relates to recombinant human interleukin-3 (hIL-3) variant or mutant proteins (muteins). These hIL-3 muteins contain amino acid substitutions and may also have amino acid deletions at both the N- and C- termini. The invention also relates to pharmaceutical compositions containing the hIL-3 muteins and methods for using them. Additionally, the present invention relates to recombinant expression vectors comprising nucleotide sequences encoding the hIL-3 muteins, related microbial expression systems, and processes for making the hIL-3 muteins using the microbial expression systems. Included in the present invention are deletion mutants of hIL-3 in which from 1 to 14 amino acids have been deleted from the N-terminus, and from 1 to 15 amino acids 119 to 133 have been deleted from the C-terminus, and which also contain amino acid substitutions in the polypeptide. These hIL-3 multiple mutation polypeptides may have biological activities similar to or better than hIL-3 and, in some cases, may also have an improved side effect profile.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . A human interleukin-3 mutant polypeptide Formula I:
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
[SEQ ID NO:15]
1 5 10 15
Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
65 70 75
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
80 85 90
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
95 100 105
Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
110 115 120
Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe
125 130
wherein
Xaa at position 17 is Ser, Lys, Gly, Asp, Met, Gln, or Arg;
Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;
Xaa at position 19 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;
Xaa at position 20 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;
Xaa at position 21 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn, Thr, Ser or Val;
Xaa at position 22 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val or Gly;
Xaa at position 23 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Leu, Ser, or Arg;
Xaa at position 24 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;
Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;
Xaa at position 26 is His, Thr, Phe, Gly, Arg, Ala, or Trp;
Xaa at position 27 is Leu, Gly, Arg, Thr, Ser, or Ala;
Xaa at position 28 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;
Xaa at position 29 is Gln, Asn, Leu, Pro, Arg, or Val;
Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys;
Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;
Xaa at position 32 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 33 is Pro, Leu, Gln, Ala, Thr, or Glu;
Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr, Arg, Ala, Phe, Ile or Met;
Xaa at position 35 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;
Xaa at position 36 is Asp, Leu, or Val;
Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;
Xaa at position 38 is Asn, or Ala;
Xaa at position 40 is Leu, Trp, or Arg;
Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, or Pro;
Xaa at position 42 is Gly, Asp, Ser, Cys, Asn, Lys, Thr, Leu, Val, Glu, Phe, Tyr, Ile, Met or Ala;
Xaa at position 43 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln, Arg, Thr, Gly or Ser;
Xaa at position 44 is Asp, Ser, Leu, Arg, Lys, Thr, Met, Trp, Glu, Asn, Gln, Ala or Pro;
Xaa at position 45 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys, Trp, Asp, Asn, Arg, Ser, Ala, Ile, Glu or His;
Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Glu, Asn, Gln, Lys, His, Ala, Tyr, Ile, Val or Gly;
Xaa at position 47 is Ile, Gly, Val, Ser, Arg, Pro, or His;
Xaa at position 48 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu, Lys, Thr, Ala, Met, Val or Asn;
Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;
Xaa at position 50 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile, Val, His, Phe, Met or Gln;
Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 52 is Asn, His, Arg, Leu, Gly, Ser, or Thr;
Xaa at position 53 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, or Met;
Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys, His, Ala or Leu;
Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly;
Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala, Tyr, Phe, Leu, Val or Lys;
Xaa at position 57 is Asn or Gly;
Xaa at position 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;
Xaa at position 59 is Glu Tyr, His, Leu, Pro, or Arg;
Xaa at position 60 is Ala, Ser, Pro, Tyr, Asn, or Thr;
Xaa at position 61 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;
Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, Asp, or Ile;
Xaa at position 63 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val;
Xaa at position 64 is Ala, Asn, Pro, Ser, or Lys;
Xaa at position 65 is Val, Thr, Pro, His, Leu, Phe, or Ser;
Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;
Xaa at position 67 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, or His;
Xaa at position 68 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His;
Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, or Leu;
Xaa at position 70 is Asn, Leu, Val, Trp, Pro, or Ala;
Xaa at position 71 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln, Trp, or Asn;
Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;
Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg;
Xaa at position 74 is Ile, Met, Thr, Pro, Arg, Gly, Ala;
Xaa at position 75 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, Gln, or Leu;
Xaa at position 76 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, or Asp;
Xaa at position 77 is Ile, Ser, Arg, Thr, or Leu;
Xaa at position 78 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;
Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile, Gly, or Asp;
Xaa at position 80 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg;
Xaa at position 81 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;
Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;
Xaa at position 83 is Pro, Ala, Thr, Trp, Arg, or Met;
Xaa at position 84 is Cys, Glu, Gly, Arg, Met, or Val;
Xaa at position 85 is Leu, Asn, Val, or Gln;
Xaa at position 86 is Pro, Cys, Arg, Ala, or Lys;
Xaa at position 87 is Leu, Ser, Trp, or Gly;
Xaa at position 88 is Ala, Lys, Arg, Val, or Trp;
Xaa at position 89 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn, or Ser;
Xaa at position 90 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or Met;
Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;
Xaa at position 92 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ile or Leu;
Xaa at position 93 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;
Xaa at position 94 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys, His, Ala, or Pro;
Xaa at position 95 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser, Ala, Trp, Phe, Ile, or Tyr;
Xaa at position 96 is Pro, Lys, Tyr, Gly, Ile, or Thr;
Xaa at position 97 is Ile, Val, Lys, Ala, or Asn;
Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;
Xaa at position 99 is Ile, Leu, Arg, Asp, Val, Pro, Gln, Gly, Ser, Phe, or His;
Xaa at position 100 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln, or Pro;
Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu, or Gln;
Xaa at position 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;
Xaa at position 103 is Asp, or Ser;
Xaa at position 104 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln, Lys, Ala, Phe, or Gly;
Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;
Xaa at position 108 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His, Ser, Ala or Pro;
Xaa at position 109 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;
Xaa at position 110 is Lys, Ala, Asn, Thr, Leu, Arg, Gln, His, Glu, Ser, Ala, or Trp;
Xaa at position 111 is Leu, Ile, Arg, Asp, or Met;
Xaa at position 112 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;
Xaa at position 113 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp, Lys, Leu, Ile, Val or Asn;
Xaa at position 114 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;
Xaa at position 115 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr, Trp, or Met;
Xaa at position 116 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu, Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;
Xaa at position 117 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;
Xaa at position 118 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;
Xaa at position 119 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;
Xaa at position 120 is Asn, Ala, Pro, Leu, His, Val, or Gln;
Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;
Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3.
2 . A human interleukin-3 mutant polypeptide of the Formula II:
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
[SEQ ID NO:16]
1 5 10 15
Cys Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa
35 40 45
Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
65 70 75
Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa Xaa
80 85 90
Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa
95 100 105
Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa Xaa
110 115 120
Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe
125 130
wherein
Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;
Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;
Xaa at position 19 is Met, Phe, Ile, Arg, or Ala;
Xaa at position 20 is Ile or Pro;
Xaa at position 21 is Asp or Glu;
Xaa at position 23 is Ile, Val, Ala, Leu, or Gly;
Xaa at position 24 is Ile, Val, Phe, or Leu;
Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;
Xaa at position 26 is His, Phe, Gly, Arg, or Ala;
Xaa at position 28 is Lys, Leu, Gln, Gly, Pro, or Val;
Xaa at position 29 is Gln, Asn, Leu, Arg, or Val;
Xaa at position 30 is Pro, His, Thr, Gly, or Gln;
Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;
Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 33 is Pro, Leu, Gln, Ala, or Glu;
Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;
Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;
Xaa at position 36 is Asp or Leu;
Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;
Xaa at position 38 is Asn or Ala;
Xaa at position 41 is Asn, Cys, Arg, His, Met, or Pro;
Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, Val or Arg;
Xaa at position 44 is Asp or Glu;
Xaa at position 45 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn, Glu, Ser, or Trp;
Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His, Ile, Lys, Tyr, Val or Gly;
Xaa at position 47 is Ile, Val, or His;
Xaa at position 49 is Met, Asn, or Asp;
Xaa at position 50 is Glu, Thr, Ala, Asn, Ser or Asp;
Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 52 is Asn or Gly;
Xaa at position 53 is Leu, Met, or Phe;
Xaa at position 54 is Arg, Ala, or Ser;
Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;
Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn, Glu, His, Leu, Thr, Val or Lys;
Xaa at position 59 is Glu, Tyr, His, Leu, or Arg;
Xaa at position 60 is Ala, Ser, Asn, or Thr;
Xaa at position 61 is Phe or Ser;
Xaa at position 62 is Asn, Val, Pro, Thr, or Ile;
Xaa at position 63 is Arg, Tyr, Lys, Ser, His, or Val;
Xaa at position 64 is Ala or Asn;
Xaa at position 65 is Val, Thr, Leu, or Ser;
Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;
Xaa at position 67 is Ser, Phe, Val, Gly, Asn, Ile, or His;
Xaa at position 68 is Leu, Val, Ile, Phe, or His;
Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;
Xaa at position 70 is Asn or Pro;
Xaa at position 71 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;
Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;
Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, or Pro;
Xaa at position 74 is Ile or Met;
Xaa at position 75 is Glu, Gly, Asp, Ser, or Gln;
Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or Asp;
Xaa at position 77 is Ile, Ser, or Leu;
Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp;
Xaa at position 80 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;
Xaa at position 81 is Leu, or Val;
Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 83 is Pro, Ala, Thr, Trp, or Met;
Xaa at position 85 is Leu or Val;
Xaa at position 87 is Leu or Ser;
Xaa at position 88 is Ala, Arg, or Trp;
Xaa at position 89 is Thr, Asp, Glu, His, Asn, or Ser;
Xaa at position 90 is Ala, Asp, or Met;
Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;
Xaa at position 92 is Pro or Ser;
Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;
Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser or Thr;
Xaa at position 96 is Pro or Tyr;
Xaa at position 97 is Ile, Val, or Ala;
Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Leu, Arg, Gln, Glu, Lys, Met, Ser, Tyr, Val or Pro;
Xaa at position 99 is Ile, Leu, Val, or Phe;
Xaa at position 100 is Lys, Leu, His, Arg, Ile, Gln, Pro, or Ser;
Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu or Tyr;
Xaa at position 102 is Gly, Glu, Lys, or Ser;
Xaa at position 104 is Trp, Val, Tyr, Met, or Leu;
Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 106 is Glu, Ser, Ala, or Gly;
Xaa at position 108 is Arg, Ala, Gln, Ser or Lys;
Xaa at position 109 is Arg, Thr, Glu, Leu, Ser, or Gly;
Xaa at position 112 is Thr, Val, Gln, Glu, His, or Ser;
Xaa at position 114 is Tyr or Trp;
Xaa at position 115 is Leu or Ala;
Xaa at position 116 is Lys, Thr, Met, Val, Trp, Ser, Leu, Ala, Asn, Gln, His, Met, Phe, Tyr or Ile;
Xaa at position 117 is Thr, Ser, or Asn;
Xaa at position 119 is Glu, Ser, Pro, Leu, Thr, or Tyr;
Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;
Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3.
3 . A human interleukin-3 mutant polypeptide according to claim 2 of the Formula III:
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
[SEQ ID NO:17]
1 5 10 15
Cys Xaa Xaa Xaa Ile Xaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa
35 40 45
Xaa Ile Leu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu Xaa
50 55 60
Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu
65 70 75
Xaa Xaa Leu Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr Ala
80 85 90
Xaa Pro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa
95 100 105
Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu Xaa
110 115 120
Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe
125 130
wherein
Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;
Xaa at position 18 is Asn, His, or Ile;
Xaa at position 19 is Met or Ile;
Xaa at position 21 is Asp or Glu;
Xaa at position 23 is Ile, Ala, Leu, or Gly;
Xaa at position 24 is Ile, Val, or Leu;
Xaa at position 25 is Thr, His, Gln, or Ala;
Xaa at position 26 is His or Ala;
Xaa at position 29 is Gln, Asn, or Val;
Xaa at position 30 is Pro, Gly, or Gln;
Xaa at position 31 is Pro, Asp, Gly, or Gln;
Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 33 is Pro or Glu;
Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;
Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;
Xaa at position 37 is Phe, Ser, Pro, or Trp;
Xaa at position 38 is Asn or Ala;
Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr or Arg;
Xaa at position 44 is Asp or Glu;
Xaa at position 45 is Gln, Val, Met, Leu, Thr, Ala, Asn, Glu, Ser or Lys;
Xaa at position 46 is Asp, Phe, Ser, Thr, Ala, Asn Gln, Glu, His, Ile, Lys, Tyr, Val or Cys;
Xaa at position 50 is Glu, Ala, Asn, Ser or Asp;
Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 54 is Arg or Ala;
Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;
Xaa at position 56 is Pro, Gly, Ser, Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val or Lys;
Xaa at position 60 is Ala or Ser;
Xaa at position 62 is Asn, Pro, Thr, or Ile;
Xaa at position 63 is Arg or Lys;
Xaa at position 64 is Ala or Asn;
Xaa at position 65 is Val or Thr;
Xaa at position 66 is Lys or Arg;
Xaa at position 67 is Ser, Phe, or His;
Xaa at position 68 is Leu, Ile, Phe, or His;
Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;
Xaa at position 71 is Ala, Pro, or Arg;
Xaa at position 72 is Ser, Glu, Arg, or Asp;
Xaa at position 73 is Ala or Leu;
Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;
Xaa at position 77 is Ile or Leu;
Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp;
Xaa at position 80 is Asn, Gly, Glu, or Arg;
Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Ile, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 83 is Pro or Thr;
Xaa at position 85 is Leu or Val;
Xaa at position 87 is Leu or Ser;
Xaa at position 88 is Ala or Trp;
Xaa at position 91 is Ala or Pro;
Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;
Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe, Ser or Thr;
Xaa at position 96 is Pro or Tyr;
Xaa at position 97 is Ile or Val;
Xaa at position 98 is His, Ile, Asn, Leu, Ala, Thr, Leu, Arg, Gln, Leu, Lys, Met, Ser, Tyr, Val or Pro;
Xaa at position 99 is Ile, Leu, or Val;
Xaa at position 100 is Lys, Arg, Ile, Gln, Pro, or Ser;
Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Pro, Asn, Ile, Leu or Tyr;
Xaa at position 104 is Trp or Leu;
Xaa at position 105 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 106 is Glu or Gly;
Xaa at position 108 is Arg, Ala, or Ser;
Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;
Xaa at position 112 is Thr, Val, or Gln;
Xaa at position 114 is Tyr or Trp;
Xaa at position 115 is Leu or Ala;
Xaa at position 116 is Lys, Thr, Val, Trp, Ser, Ala, His, Met, Phe, Tyr or Ile;
Xaa at position 117 is Thr or Ser;
Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;
Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 35 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3.
4 . A human interieukin-3 mutant polypeptide according to claim 3 of the Formula IV:
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
[SEQ ID NO:18]
1 5 10 15
Cys Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa Xaa
20 25 30
Pro Xaa Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp Xaa
35 40 45
Xaa Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu Ala
50 55 60
Phe Xaa Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile Glu
65 70 75
Xaa Xaa Leu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr Ala
80 85 90
Xaa Pro Xaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp Xaa
95 100 105
Glu Phe Xaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu Xaa
110 115 120
Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe
125 130
wherein
Xaa at position 17 is Ser, Gly, Asp, or Gln;
Xaa at position 18 is Asn, His, or Ile;
Xaa at position 23 is Ile, Ala, Leu, or Gly;
Xaa at position 25 is Thr, His, or Gln;
Xaa at position 26 is His or Ala;
Xaa at position 29 is Gln or Asn;
Xaa at position 30 is Pro or Gly;
Xaa at position 32 is Leu, Arg, Asn, or Ala;
Xaa at position 34 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr, or Met;
Xaa at position 35 is Leu, Ala, Asn, or Pro;
Xaa at position 38 is Asn or Ala;
Xaa at position 42 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr or Arg;
Xaa at position 45 is Gln, Val, Met, Leu, Ala, Asn, Glu, or Lys;
Xaa at position 46 is Asp, Phe, Ser, Gln, Glu, His, Val or Thr;
Xaa at position 50 is Glu Asn, Ser or Asp;
Xaa at position 51 is Asn, Arg, Pro, Thr, or His;
Xaa at position 55 is Arg, Leu, or Gly;
Xaa at position 56 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;
Xaa at position 62 is Asn, Pro, or Thr;
Xaa at position 64 is Ala or Asn;
Xaa at position 65 is Val or Thr;
Xaa at position 67 is Ser or Phe;
Xaa at position 68 is Leu or Phe;
Xaa at position 69 is Gln, Ala, Glu, or Arg;
Xaa at position 76 is Ser, Val, Asn, Pro, or Gly;
Xaa at position 77 is Ile or Leu;
Xaa at position 79 is Lys, Gly, Asn, Met, Arg, Ile, or Gly;
Xaa at position 80 is Asn, Gly, Glu, or Arg;
Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 87 is Leu or Ser;
Xaa at position 88 is Ala or Trp;
Xaa at position 91 is Ala or Pro;
Xaa at position 93 is Thr, Asp, or Ala;
Xaa at position 95 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;
Xaa at position 98 is His, Ile, Asn, Ala, Thr, Gln, Glu, Lys, Met, Ser, Tyr, Val or Leu;
Xaa at position 99 is Ile or Leu;
Xaa at position 100 is Lys or Arg;
Xaa at position 101 is Asp, Pro, Met, Lys, Thr, His, Pro, Asn, Ile, Leu or Tyr;
Xaa at position 105 is Asn, Pro, Ser, Ile or Asp;
Xaa at position 108 is Arg, Ala, or Ser;
Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;
Xaa at position 112 is Thr or Gln;
Xaa at position 116 is Lys, Val, Trp, Ala, His, Phe, Tyr or Ile;
Xaa at position 117 is Thr or Ser;
Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 121 is Ala, Ser, Ile, Pro, or Asp;
Xaa at position 122 is Gln, Met, Trp, Phe, Pro, His, Ile, or Tyr;
Xaa at position 123 is Ala, Met, Glu, Ser, or Leu;
and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3.
5 . The human interleukin-3 mutant polypeptide of claim 1 wherein 1-15 amino acids are deleted from the C-terminus and/or 1-14 amino acids are deleted from the N-terminus.
6 . The human interleukin-3 mutant polypeptide of claim 1 wherein;
Xaa at position 42 is Gly, Asp, Ser, Ile, Leu, Met, Tyr, or Ala;
Xaa at position 45 is Gln, Val, Met or Asn;
Xaa at position 46 is Asp, Ser, Gln, His or Val;
Xaa at position 50 is Glu or Asp;
Xaa at position 51 is Asn, Pro or Thr;
Xaa at position 62 is Asn or Pro;
Xaa at position 76 is Ser, or Pro;
Xaa at position 82 is Leu, Trp, Asp, Asn Glu, His, Phe, Ser or Tyr;
Xaa at position 95 is His, Arg, Thr, Asn or Ser;
Xaa at position 98 is His, Ile, Leu, Ala, Gln, Lys, Met, Ser, Tyr or Val;
Xaa at position 100 is Lys or Arg;
Xaa at position 101 is Asp, Pro, His, Asn, Ile or Leu;
Xaa at position 105 is Asn, or Pro;
Xaa at position 108 is Arg, Ala, or Ser;
Xaa at position 116 is Lys, Val, Trp, Ala, His, Phe, or Tyr;
Xaa at position 121 is Ala, or Ile;
Xaa at position 122 is Gln, or Ile; and
Xaa at position 123 is Ala, Met or Glu.
7 . A (15-125) human interleukin-3 mutant polypeptide of the Formula V:
Asn Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
[SEQ ID NO:19]
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
65 70 75
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
80 85 90
Xaa Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
95 100 105
Xaa Xaa Xaa Xaa Gln Gln
110
wherein
Xaa at position 3 is Ser, Lys, Gly, Asp, Met, Gln, or Arg;
Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;
Xaa at position 5 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;
Xaa at position 6 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;
Xaa at position 7 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn, Thr, Ser or Val;
Xaa at position 8 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val, or Gly;
Xaa at position 9 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Leu, Ser, or Arg;
Xaa at position 10 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;
Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;
Xaa at position 12 is His, Thr, Phe, Gly, Arg, Ala, or Trp;
Xaa at position 13 is Leu, Gly, Arg, Thr, Ser, or Ala;
Xaa at position 14 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;
Xaa at position 15 is Gln, Asn, Leu, Pro, Arg, or Val;
Xaa at position 16 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys;
Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;
Xaa at position 18 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 19 is Pro, Leu, Gln, Ala, Thr, or Glu;
Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr, Arg, Ala, Phe, Ile or Met;
Xaa at position 21 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;
Xaa at position 22 is Asp, Leu, or Val;
Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;
Xaa at position 24 is Asn, or Ala;
Xaa at position 26 is Leu, Trp, or Arg;
Xaa at position 27 is Asn, Cys, Arg, Leu, His, Met, Pro;
Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Lys, Asn, Thr, Leu, Val, Glu, Phe, Tyr, Ile or Met;
Xaa at position 29 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln, Arg, Thr, Gly or Ser;
Xaa at position 30 is Asp, Ser, Leu, Arg, Lys, Thr,Met, Trp, Glu, Asn, Gln, Ala or Pro;
Xaa at position 31 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys, Asp, Asn, Arg, Ser, Ala, Ile, Glu, His or Trp;
Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Glu, Asn, Gln, Lys, His, Ala, Tyr, Ile, Val or Gly;
Xaa at position 33 is Ile, Gly, Val, Ser, Arg, Pro, or His;
Xaa at position 34 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu, Lys, Thr, Ala, Met, Val or Asn;
Xaa at position 35 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;
Xaa at position 36 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile, Val, His, Phe, Met or Gln;
Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 38 is Asn, His, Arg, Leu, Gly, Ser, or Thr;
Xaa at position 39 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, Met, or;
Xaa at position 40 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys, His, Ala or Leu;
Xaa at position 41 is Arg, Thr, Val, Ser, Leu, or Gly;
Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala, Tyr, Phe, Leu, Val or Lys;
Xaa at position 43 is Asn or Gly;
Xaa at position 44 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;
Xaa at position 45 is Glu Tyr, His, Leu, Pro, or Arg;
Xaa at position 46 is Ala, Ser, Pro, Tyr, Asn, or Thr;
Xaa at position 47 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;
Xaa at position 48 is Asn, His, Val, Arg, Pro, Thr, Asp, or Ile;
Xaa at position 49 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val;
Xaa at position 50 is Ala, Asn, Pro, Ser, or Lys;
Xaa at position 51 is Val, Thr, Pro, His, Leu, Phe, or Ser;
Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;
Xaa at position 53 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, or His;
Xaa at position 54 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His;
Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, or Leu;
Xaa at position 56 is Asn, Leu, Val, Trp, Pro, or Ala;
Xaa at position 57 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln, Trp, or Asn;
Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;
Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg;
Xaa at position 60 is Ile, Met, Thr, Pro, Arg, Gly, Ala;
Xaa at position 61 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, Gln, or Leu;
Xaa at position 62 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, or Asp;
Xaa at position 63 is Ile, Ser, Arg, Thr, or Leu;
Xaa at position 64 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;
Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or Asp;
Xaa at position 66 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg;
Xaa at position 67 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;
Xaa at position 68 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;
Xaa at position 69 is Pro, Ala, Thr, Trp, Arg, or Met;
Xaa at position 70 is Cys, Glu, Gly, Arg, Met, or Val;
Xaa at position 71 is Leu, Asn, Val, or Gln;
Xaa at position 72 is Pro, Cys, Arg, Ala, or Lys;
Xaa at position 73 is Leu, Ser, Trp, or Gly;
Xaa at position 74 is Ala, Lys, Arg, Val, or Trp;
Xaa at position 75 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn, or Ser;
Xaa at position 76 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or Met;
Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;
Xaa at position 78 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ile or Leu;
Xaa at position 79 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;
Xaa at position 80 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys, His, Ala or Pro;
Xaa at position 81 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser, Ala, Trp, Phe, Ile or Tyr;
Xaa at position 82 is Pro, Lys, Tyr, Gly, Ile, or Thr;
Xaa at position 83 is Ile, Val, Lys, Ala, or Asn;
Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;
Xaa at position 85 is Ile, Leu, Arg, Asp, Val, Pro, Gln, Gly, Ser, Phe, or His;
Xaa at position 86 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln, Pro;
Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu or Gln;
Xaa at position 88 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;
Xaa at position 89 is Asp, or Ser;
Xaa at position 90 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln, Lys, Ala, Phe, or Gly;
Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 92 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;
Xaa at position 94 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His, Ser, Ala, or Pro;
Xaa at position 95 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;
Xaa at position 96 is Lys, Asn, Thr, Leu, Gln, Arg, His, Glu, Ser, Ala or Trp;
Xaa at position 97 is Leu, Ile, Arg, Asp, or Met;
Xaa at position 98 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;
Xaa at position 99 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp, Lys, Leu, Ile, Val or Asn;
Xaa at position 100 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;
Xaa at position 101 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr, Trp, or Met;
Xaa at position 102 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu, Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;
Xaa at position 103 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;
Xaa at position 104 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;
Xaa at position 105 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;
Xaa at position 106 is Asn, Ala, Pro, Leu, His, Val, or Gln;
Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;
Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding native amino acids of (1-133) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.
8 . A (15-125) human interleukin-3 mutant polypeptide of the Formula VI:
Asn Cys Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa
[SEQ ID NO:20]
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa
20 25 30
Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa
35 40 45
Xaa Xaa Xea Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa
65 70 75
Xaa Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa
80 85 90
Xaa Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa
95 100 105
Xaa Xaa Xaa Xaa Gln Gln
110
wherein
Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;
Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;
Xaa at position 5 is Met, Phe, Ile, Arg, or Ala;
Xaa at position 6 is Ile or Pro;
Xaa at position 7 is Asp, or Glu;
Xaa at position 9 is Ile, Val, Ala, Leu, or Gly;
Xaa at position 10 is Ile, Val, Phe, or Leu;
Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;
Xaa at position 12 is His, Phe, Gly, Arg, or Ala;
Xaa at position 14 is Lys, Leu, Gln, Gly, Pro, or Val;
Xaa at position 15 is Gln, Asn, Leu, Arg, or Val;
Xaa at position 16 is Pro, His, Thr, Gly, or Gln;
Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;
Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 19 is Pro, Leu, Gln, Ala, or Glu;
Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;
Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;
Xaa at position 22 is Asp or Leu;
Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;
Xaa at position 24 is Asn or Ala;
Xaa at position 27 is Asn, Cys, Arg, His, Met, or Pro;
Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, or Arg;
Xaa at position 30 is Asp, or Glu;
Xaa at position 31 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn Glu, Ser or Trp;
Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His, Ile, Lys, Tyr, Val or Gly;
Xaa at position 33 is Ile, Val, or His;
Xaa at position 35 is Met, Asn, or Asp;
Xaa at position 36 is Glu, Thr, Ala, Asn, Ser or Asp;
Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 38 is Asn or Gly;
Xaa at position 39 is Leu, Met, or Phe;
Xaa at position 40 is Arg, Ala or Ser;
Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;
Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn, Glu, His, Leu, Thr, Val or Lys;
Xaa at position 45 is Glu, Tyr, His, Leu, or Arg;
Xaa at position 46 is Ala, Ser, Asn, or Thr;
Xaa at position 47 is Phe or Ser;
Xaa at position 48 is Asn, Val, Pro, Thr, or Ile;
Xaa at position 49 is Arg, Tyr, Lys, Ser, His, or Val;
Xaa at position 50 is Ala or Asn;
Xaa at position 51 is Val, Thr, Leu, or Ser;
Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;
Xaa at position 53 is Ser, Phe, Val, Gly, Asn, Ile, or His;
Xaa at position 54 is Leu, Val, Ile, Phe, or His;
Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;
Xaa at position 56 is Asn or Pro;
Xaa at position 57 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;
Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;
Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, or Pro;
Xaa at position 60 is Ile or Met;
Xaa at position 61 is Glu, Gly, Asp, Ser, or Gln;
Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or Asp;
Xaa at position 63 is Ile, Ser, or Leu;
Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or Asp;
Xaa at position 66 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;
Xaa at position 67 is Leu, or Val;
Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 69 is Pro, Ala, Thr, Trp, or Met;
Xaa at position 71 is Leu or Val;
Xaa at position 73 is Leu or Ser;
Xaa at position 74 is Ala, Arg, or Trp;
Xaa at position 75 is Thr, Asp, Glu, His, Asn, or Ser;
Xaa at position 76 is Ala, Asp, or Met;
Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;
Xaa at position 78 is Pro or Ser;
Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;
Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser or Thr;
Xaa at position 82 is Pro or Tyr;
Xaa at position 83 is Ile, Val, or Ala;
Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr, Arg, Gln, Glu, Lys, Met, Ser, Tyr, Val or Pro;
Xaa at position 85 is Ile, Leu, Val, or Phe;
Xaa at position 86 is Lys, Leu, His, Arg, Ile, Gln, Pro or Ser;
Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu or Tyr;
Xaa at position 88 is Gly, Glu, Lys, or Ser;
Xaa at position 90 is Trp, Val, Tyr, Met, or Leu;
Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 92 is Glu, Ser, Ala, or Gly;
Xaa at position 94 is Arg, Ala, Gln, Ser or Lys;
Xaa at position 95 is Arg, Thr, Glu, Leu, Ser, or Gly;
Xaa at position 98 is Thr, Val, Gln, Glu, His, or Ser;
Xaa at position 100 is Tyr or Trp;
Xaa at position 101 is Leu or Ala;
Xaa at position 102 is Lys, Thr, Met, Val, Trp, Ser, Leu, Ala, Asn, Gln, His, Met, Phe, Tyr or Ile;
Xaa at position 103 is Thr, Ser, or Asn;
Xaa at position 105 is Glu, Ser, Pro, Leu, Thr, or Tyr;
Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;
Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.
9 . A (15-125) human interleukin-3 mutant polypeptide according to claim 7 of the Formula VII:
Asn Cys Xaa Xaa Xaa Ile Xaa Gln Xaa Xaa Xaa Xaa Leu Lys Xaa
[SEQ ID NO:21]
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa
20 25 30
Xaa Xaa Ile Leu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Gln
35 40 45
Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile
50 55 60
Gln Xaa Xaa Leu Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr
65 70 75
Ala Xaa Pro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa
80 85 90
Xaa Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu
95 100 105
Xaa Xaa Xaa Xaa Gln Gln
110
wherein
Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;
Xaa at position 4 is Asn, His, or Ile;
Xaa at position 5 is Met or Ile;
Xaa at position 7 is Asp or Glu;
Xaa at position 9 is Ile, Ala, Leu, or Gly;
Xaa at position 10 is Ile, Val, or Leu;
Xaa at position 11 is Thr, His, Gln, or Ala;
Xaa at position 12 is His or Ala;
Xaa at position 15 is Gln, Asn, or Val;
Xaa at position 16 is Pro, Gly, or Gln;
Xaa at position 17 is Pro, Asp, Gly, or Gln;
Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 19 is Pro or Glu;
Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;
Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;
Xaa at position 23 is Phe, Ser, Pro, or Trp;
Xaa at position 24 is Asn or Ala;
Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met Tyr or Arg;
Xaa at position 30 is Asp or Glu;
Xaa at position 31 is Gln, Val, Met, Leu, Thr, Ala, Asn, Glu, Ser or Lys;
Xaa at position 32 is Asp, Phe, Ser, Thr, Ala, Asn, Gln, Glu, His, Ile, Lys, Tyr, Val or Cys;
Xaa at position 36 is Glu, Ala, Asn, Ser or Asp;
Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 40 is Arg or Ala;
Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;
Xaa at position 42 is Pro, Gly, Ser, Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val or Lys;
Xaa at position 46 is Ala or Ser;
Xaa at position 48 is Asn, Pro, Thr, or Ile;
Xaa at position 49 is Arg or Lys;
Xaa at position 50 is Ala or Asn;
Xaa at position 51 is Val or Thr;
Xaa at position 52 is Lys or Arg;
Xaa at position 53 is Ser, Phe, or His;
Xaa at position 54 is Leu, Ile, Phe, or His;
Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;
Xaa at position 57 is Ala, Pro, or Arg;
Xaa at position 58 is Ser, Glu, Arg, or Asp;
Xaa at position 59 is Ala or Leu;
Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;
Xaa at position 63 is Ile or Leu;
Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp;
Xaa at position 66 is Asn, Gly, Glu, or Arg;
Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Ile, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 69 is Pro or Thr;
Xaa at position 71 is Leu or Val;
Xaa at position 73 is Leu or Ser;
Xaa at position 74 is Ala or Trp;
Xaa at position 77 is Ala or Pro;
Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;
Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe, Ser or Thr;
Xaa at position 82 is Pro or Tyr;
Xaa at position 83 is Ile or Val;
Xaa at position 84 is His, Ile, Asn, Leu, Ala, Thr, Leu, Arg, Gln, Leu, Lys, Met, Ser, Tyr, Val or Pro;
Xaa at position 85 is Ile, Leu, or Val;
Xaa at position 86 is Lys, Arg, Ile, Gln, Pro, or Ser;
Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Asn, Ile, Leu or Tyr;
Xaa at position 90 is Trp or Leu;
Xaa at position 91 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His;
Xaa at position 92 is Glu, or Gly;
Xaa at position 94 is Arg, Ala, or Ser;
Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;
Xaa at position 98 is Thr, Val, or Gln;
Xaa at position 100 is Tyr or Trp;
Xaa at position 101 is Leu or Ala;
Xaa at position 102 is Lys, Thr, Val, Trp, Ser, Ala, His, Met, Phe, Tyr or Ile;
Xaa at position 103 is Thr or Ser;
Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;
Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys;
Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;
which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 35 of the amino acids designated by Xaa are different from the corresponding amino acids of native human interleukin-3.
10 . A (15-125) human interieukin-3 mutant polypeptide according to claim 7 of the Formula VIII:
Asn Cys Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa
[SEQ ID NO:22]
1 5 10 15
Xaa Pro Xaa Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Gln Asp
20 25 30
Xaa Xaa Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Gln
35 40 45
Ala Phe Xaa Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile
50 55 60
Glu Xaa Xaa Leu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr
65 70 75
Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp
80 85 90
Xaa Gln Phe Xaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Gln
95 100 105
Xaa Xaa Xaa Xaa Gln Gln
110
wherein
Xaa at position 3 is Ser, Gly, Asp, or Gln;
Xaa at position 4 is Asn, His, or Ile;
Xaa at position 9 is Ile, Ala, Leu, or Gly;
Xaa at position 11 is Thr, His, or Gln;
Xaa at position 12 is His or Ala;
Xaa at position 15 is Gln or Asn;
Xaa at position 16 is Pro or Gly;
Xaa at position 18 is Leu, Arg, Asn, or Ala;
Xaa at position 20 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;
Xaa at position 21 is Leu, Ala, Asn, or Pro;
Xaa at position 24 is Asn or Ala;
Xaa at position 28 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr or Arg;
Xaa at position 31 is Gln, Val, Met, Leu, Ala, Asn, Glu or Lys;
Xaa at position 32 is Asp, Phe, Ser, Ala, Gln, Glu, His, Val or Thr;
Xaa at position 36 is Gln, Asn, Ser or Asp;
Xaa at position 37 is Asn, Arg, Pro, Thr, or His;
Xaa at position 41 is Arg, Leu, or Gly;
Xaa at position 42 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;
Xaa at position 48 is Asn, Pro, or Thr;
Xaa at position 50 is Ala or Asn;
Xaa at position 51 is Val or Thr;
Xaa at position 53 is Ser or Phe;
Xaa at position 54 is Leu or Phe;
Xaa at position 55 is Gln, Ala, Glu, or Arg;
Xaa at position 62 is Ser, Val, Asn, Pro, or Gly;
Xaa at position 63 is Ile or Leu;
Xaa at position 65 is Lys, Asn, Met, Arg, Ile, or Gly;
Xaa at position 66 is Asn, Gly, Glu, or Arg;
Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val;
Xaa at position 73 is Leu or Ser;
Xaa at position 74 is Ala or Trp;
Xaa at position 77 is Ala or Pro;
Xaa at position 79 is Thr, Asp, or Ala;
Xaa at position 81 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;
Xaa at position 84 is His, Ile, Asn, Ala, Thr, Arg, Gln, Glu, Lys, Met, Ser, Tyr, Val or Leu;
Xaa at position 85 is Ile or Leu;
Xaa at position 86 is Lys or Arg;
Xaa at position 87 is Asp, Pro, Met, Lys, His, Pro, Asn, Ile, Leu or Tyr;
Xaa at position 91 is Asn, Pro, Ser, Ile or Asp;
Xaa at position 94 is Arg, Ala, or Ser;
Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;
Xaa at position 98 is Thr or Gln;
Xaa at position 102 is Lys, Val, Trp, or Ile;
Xaa at position 103 is Thr, Ala, His, Phe, Tyr or Ser;
Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;
Xaa at position 107 is Ala, Ser, Ile, Pro, or Asp;
Xaa at position 108 is Gln, Met, Trp, Phe, Pro, His, Ile, or Tyr;
Xaa at position 109 is Ala, Met, Glu, Ser, or Leu;
and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 26 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.
11 . A (15-125) human interleukin-3 mutant polypeptide of claim 7 wherein:
Xaa at position 17 is Ser, Lys, Asp, Met, Gln, or Arg;
Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;
Xaa at position 19 is Met, Arg, Gly, Ala, or Cys;
Xaa at position 20 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;
Xaa at position 21 is Asp, Phe, Lys, Arg, Ala, Gly, or Val;
Xaa at position 22 is Glu, Trp, Pro, Ser, Ala, His, or Gly;
Xaa at position 23 is Ile, Ala, Gly, Trp, Lys, Leu, Ser, or Arg;
Xaa at position 24 is Ile, Gly, Arg, or Ser;
Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;
Xaa at position 26 is His, Thr, Phe, Gly, Ala, or Trp;
Xaa at position 27 is Leu, Gly, Arg, Thr, Ser, or Ala;
Xaa at position 28 is Lys, Leu, Gln, Gly, Pro, Val or Trp;
Xaa at position 29 is Gln, Asn, Loh, Pro, Arg, or Val;
Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys;
Xaa at position 31 is Pro, Asp, Gly, Arg, Leu, or Gln;
Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;
Xaa at position 33 is Pro, Leu, Gln, Thr, or Glu;
Xaa at position 34 is Leu, Gly, Ser, or Lys;
Xaa at position 35 is Leu, Ala, Gly, Asn, Pro, or Gln;
Xaa at position 36 is Asp, Leu, or Val;
Xaa at position 37 is Phe, Ser, or Pro;
Xaa at position 38 is Asn, or Ala;
Xaa at position 40 is Leu, Trp, or Arg;
Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, Pro;
Xaa at position 42 is Gly, Asp, Ser, Cys, or Ala;
Xaa at position 42 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys, or Ser;
Xaa at position 44 is Asp, Ser, Leu, Arg, Lys, Thr, Met, Trp, or Pro;
Xaa at position 45 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys, or Trp;
Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, or Gly;
Xaa at position 47 is Ile, Gly, Ser, Arg, Pro, or His;
Xaa at position 48 is Leu, Ser, Cys, Arg, His, Phe, or Asn;
Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;
Xaa at position 50 is Glu, Leu, Thr, Asp, or Tyr;
Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;
Xaa at position 52 is Asn, His, Arg, Leu, Gly, Ser, or Thr;
Xaa at position 53 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, or;
Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr, Gln, or Leu;
Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly;
Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, or Lys;
Xaa at position 57 is Asn or Gly;
Xaa at position 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;
Xaa at position 59 is Glu Tyr, His, Leu, Pro, or Arg;
Xaa at position 60 is Ala, Ser, Tyr, Asn, or Thr;
Xaa at position 61 is Phe, Asn, Gln, Pro, Lys, Arg, or Ser;
Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, or Ile;
Xaa at position 63 is Arg, Tyr, Trp, Ser, Pro, or Val;
Xaa at position 64 is Ala, Asn, Ser, or Lys;
Xaa at position 65 is Val, Thr, Pro, His, Leu, Phe, or Ser;
Xaa at position 66 is Lys, Ile, Val, Asn, Gln, or Ser;
Xaa at position 67 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, or His;
Xaa at position 68 is Leu, Val, Trp, Ser, Thr, or His;
Xaa at position 69 is Gln, Ala, Pro, Thr, Arg, Trp, Gly, or Leu;
Xaa at position 70 is Asn, Leu, Val, Trp, Pro, or Ala;
Xaa at position 71 is Ala, Met, Leu, Arg, Glu, Thr, Gln, Trp, or Asn;
Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;
Xaa at position 73 is Ala, Gln, Asp, Leu, Ser, Gly, Thr, or Arg;
Xaa at position 74 is Ile, Thr, Pro, Arg, Gly, Ala;
Xaa at position 75 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, or Leu;
Xaa at position 76 is Ser, Val, Ala, Asn, Trp, Gln, Pro, Gly, or Asp;
Xaa at position 77 is Ile, Ser, Arg, or Thr;
Xaa at position 78 is Leu, Ala, Ser, Glu, Gly, or Arg;
Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Ile, or Asp;
Xaa at position 80 is Asn, Trp, Val, Gly, Thr, Leu, or Arg;
Xaa at position 81 is Leu, Gln, Gly, Ala, Trp, Arg, or Lys;
Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, or Asp;
Xaa at position 83 is Pro, Thr, Trp, Arg, or Met;
Xaa at position 84 is Cys, Gln, Gly, Arg, Met, or Val;
Xaa at position 85 is Leu, Asn, or Gln;
Xaa at position 86 is Pro, Cys, Arg, Ala, or Lys;
Xaa at position 87 is Leu, Ser, Trp, or Gly;
Xaa at position 88 is Ala, Lys, Arg, Val, or Trp;
Xaa at position 89 is Thr, Asp, Cys, Leu, Val, Glu, His, or Asn;
Xaa at position 90 is Ala, Ser, Asp, Ile, or Met;
Xaa at position 91 is Ala, Ser, Thr, Phe, Leu, Asp, or His;
Xaa at position 92 is Pro, Phe, Arg, Ser, Lys, His, or Leu;
Xaa at position 93 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;
Xaa at position 94 is Arg, Ile, Ser, Glu, Leu, Val, or Pro;
Xaa at position 95 is His, Gln, Pro, Val, Leu, Thr or Tyr;
Xaa at position 96 is Pro, Lys, Tyr, Gly, Ile, or Thr;
Xaa at position 97 is Ile, Lys, Ala, or Asn;
Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, or Pro;
Xaa at position 99 is Ile, Arg, Asp, Pro, Gln, Gly, Phe, or His;
Xaa at position 100 is Lys, Tyr, Leu, His, Ile, Ser, Gln, or Pro;
Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr, or Gln;
Xaa at position 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;
Xaa at position 103 is Asp, or Ser;
Xaa at position 104 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln, Lys, Ala, Phe, or Gly;
Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, or His;
Xaa at position 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;
Xaa at position 108 is Arg, Asp, Leu, Thr, Ile, or Pro;
Xaa at position 109 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly.
12 . The human interleukin-3 mutant polypetide of claim 7:
wherein;
Xaa at position 28 is Gly, Asp, Ser, Ile, Leu, Met, Tyr, or Ala;
Xaa at position 31 is Gln, Val, Met or Asn;
Xaa at position 32 is Asp, Ser, Ala, Gln, His or Val;
Xaa at position 36 is Glu or Asp;
Xaa at position 37 is Asn, Pro or Thr;
Xaa at position 48 is Asn or Pro;
Xaa at position 62 is Ser, or Pro;
Xaa at position 68 is Leu, Trp, Asp, Asn Glu, His, Phe, Ser or Tyr;
Xaa at position 81 is His, Arg, Thr, Asn or Ser;
Xaa at position 84 is His, Ile, Leu, Ala, Arg, Gln, Lys, Met, Ser, Tyr or Val;
Xaa at position 86 is Lys or Arg;
Xaa at position 87 is Asp, Pro, His, Asn, Ile or Leu;
Xaa at position 91 is Asn, or Pro;
Xaa at position 94 is Arg, Ala, or Ser;
Xaa at position 102 is Lys, Val, Trp, Ala, His, Phe, or Tyr;
Xaa at position 107 is Ala, or Ile;
Xaa at position 108 is Gln, or Ile; and
Xaa at position 109 is Ala, Met or Glu.
13 . A polypeptide of the formula
1 5 10
(Met) m -Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr
[SEQ ID NO:29]
15 20
Ser Trp Val Asn Cys Ser Xaa Xaa Xaa Asp Gln Ile Ile
25 30 35
Xaa His Leu Lys Xaa Pro Pro Xaa Pro Xaa Leu Asp Xaa
40 45 50
Xaa Asn Leu Asn Xaa Gln Asp Xaa Asp Ile Leu Xaa Glu
55 60
Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa
65 70 75
Ala Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Gln Xaa
80 85
Ile Leu Xaa Asn Leu Xaa Pro Cys Xaa Pro Xaa Xaa Thr
90 95 100
Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Ile Xaa Xaa Gly
105 110 115
Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Xaa Phe Tyr Leu
120 125
Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln Thr Thr Leu
130
Ser Leu Ala Ile Phe
wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa at position 19 is Met, Ala or Ile; Xaa at position 20 is Ile, Pro or Ile; Xaa at position 23 is Ile, Ala or Leu; Xaa at position 25 is Thr or His; Xaa at position 29 is Gln, Arg, Val or Ile; Xaa at position 32 is Leu, Ala, Asn or Arg; Xaa at position 34 is Leu or Ser; Xaa at position 37 is Phe, Pro, or Ser; Xaa at position 38 is Asn or Ala; Xaa at position 42 is Gly, Ala, Ser, Asp or Asn; Xaa at position 45 is Gln, Val, or Met; Xaa at position 46 is Asp or Ser; Xaa at position 49 is Met, Ile, Leu or Asp; Xaa at position 50 is Glu or Asp; Xaa at position 51 is Asn Arg or Ser; Xaa at position 55 is Arg, Leu, or Thr; Xaa at position 56 is Pro or Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa at position 62 is Asn, Val or Pro; Xaa at position 63 is Arg or His; Xaa at position 65 is Val or Ser; Xaa at position 67 is Ser, Asn, His or Gln; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Ala or Gly; Xaa at position 76 is Ser, Ala or Pro; Xaa at position 79 is Lys, Arg or Ser; Xaa at position 82 is Leu, Glu, Val or Trp; Xaa at position 85 is Leu or Val; Xaa at position 87 is Leu, Ser, Tyr; Xaa at position 88 is Ala or Trp; Xaa at position 91 is Ala or Pro; Xaa at position 93 is Pro or Ser; Xaa at position 95 is His or Thr; Xaa at position 98 is His, Ile, or Thr; Xaa at position 100 is Lys or Arg; Xaa at position 101 is Asp, Ala or Met; Xaa at position 105 is Asn or Glu; Xaa at position 109 is Arg, Glu or Leu; Xaa at position 112 is Thr or Gln; Xaa at position 116 is Lys, Val, Trp or Ser; Xaa at position 117 is Thr or Ser; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123 is Ala or Glu; with the proviso that from four to forty-four of the amino acids designated by Xaa are different from the corresponding amino acids of native human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.
14 . A polypeptide according to claim 13 wherein Xaa at position 18 is Ile; Xaa at position 19 is Ala, or Ile; Xaa at position 20 is Pro, or Leu; Xaa at position 23 is Ala, or Leu; Xaa at position 25 is His; Xaa at position 29 is Arg, Val, or Ile; Xaa at position 32 is Ala, Asn or Arg; Xaa at position 34 is Ser; Xaa at position 37 is Pro or Ser; Xaa at position 38 is Ala; Xaa at position 42 is Ala, Ser, Asp, or Asn; and Xaa at position 45 is Val or Met; Xaa at position 46 is Ser.
15 . A polypeptide according to claim 13 wherein Xaa at position 49 is Ile, or Leu, or Asp; Xaa at position 50 is Asp; Xaa at position 51 is Arg or Ser; Xaa at position 55 is Leu or Thr; Xaa at position 56 is Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa at position 62 is Val, or Pro; Xaa at position 63 is His; Xaa at position 65 is Ser; Xaa at position 67 is Asn, or His, or Gln; and Xaa at position 69 is Glu.
16 . A polypeptide according to claim 13 wherein Xaa at position 73 is Gly; Xaa at position 76 is Ala, or Pro; Xaa at position 79 is Arg, or Ser; Xaa at position 82 is Gln or Val, or Trp; Xaa at position 85 is Val; Xaa at position 87 is Ser, or Tyr; Xaa at position 88 is Trp; Xaa at position 91 is Pro; Xaa at position 93 is Ser; Xaa at position 95 is Thr; Xaa at position 98 is Ile or Thr; Xaa at position 100 is Arg; Xaa at position 101 is Ala, or Met; and Xaa at position 105 is Glu.
17 . A polypeptide according to claim 13 wherein Xaa at position 109 is Glu, or Leu; Xaa at position 112 is Gln; Xaa at position 116 is Val, or Trp, or Ser; Xaa at position 117 is Ser; Xaa at position 120 is Glu or His; and Xaa at position 123 is Glu.
18 . A polypeptide according to claim 13 wherein Xaa at position 18 is Ile; Xaa at position 19 is Ala, or Ile; Xaa at position 20 is Pro, or Leu; Xaa at position 23 is Ala, or Leu; Xaa at position 25 is His; Xaa at position 29 is Arg or Val, or Ile; Xaa at position 32 is Ala or Asn, or Arg; Xaa at position 34 is Ser; Xaa at position 37 is Pro or Ser; Xaa at position 38 is Ala; Xaa at position 42 is Ala or Ser, Asp or Asn; Xaa at position 45 is Val or Met; Xaa at position 46 is Ser; Xaa at position 49 is Ile, or Leu, or Asp; Xaa at position 50 is Asp; Xaa at position 51 is Arg, or Ser; Xaa at position 55 is Leu or Thr; Xaa at position 56 is Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa at position 62 is Val, or Pro; Xaa at position 63 is His; Xaa at position 65 is Ser; Xaa at position 67 is Asn, or His, or Gln; and Xaa at position 69 is Glu.
19 . A polypeptide according to claim 13 wherein Xaa at position 73 is Gly; Xaa at position 76 is Ala, or Pro; Xaa at position 79 is Arg, or Ser; Xaa at position 82 is Gln or Val, or Trp; Xaa at position 85 is Val; Xaa at position 87 is Ser, or Tyr; Xaa at position 88 is Trp; Xaa at position 91 is Pro; Xaa at position 93 is Ser; Xaa at position 95 is Thr; Xaa at position 98 is Ile or Thr; Xaa at position 100 is Arg; Xaa at position 101 is Ala, or Met; Xaa at position 105 is Glu; Xaa at position 109 is Glu, or Leu; Xaa at position 112 is Gln; Xaa at position 116 is Val, or Trp, or Ser; Xaa at position 117 is Ser; Xaa at position 120 is Glu or His; and Xaa at position 123 is Glu.
20 . A polypeptide of the formula
1 5 10
(Met m -Ala n ) p Asn Cys Ser Xaa Xaa Xaa Asp Glu Xaa Ile
[SEQ ID NO: 130]
15 20
Xaa His Leu Lys Xaa Pro Pro Xaa Pro Xaa Leu Asp Xaa
25 30 35
Xaa Asn Leu Asn Xaa Gln Asp Xaa Xaa Ile Leu Xaa Gln
40 45
Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa
50 55 60
Ala Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa
65 70 75
Ile Leu Xaa Asn Xaa Xaa Pro Cys Xaa Pro Xaa Ala Thr
80 85
Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Ile Xaa Xaa Gly
90 95 100
Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Xaa Phe Tyr Leu
105 110
Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln
wherein m is 0 or 1; n is 0 or 1; p is 0 or 1; Xaa at position 4 is Asn or Ile; Xaa at position 5 is Met, Ala or Ile: Xaa at position 6 is Ile, Pro or Leu; Xaa at position 9 is Ile, Ala or Leu; Xaa at position 11 is Thr or His; Xaa at position 15 is Gln, Arg, Val or Ile; Xaa at position 18 is Leu, Ala, Asn or Arg; Xaa at position 20 is Leu or Ser; Xaa at position 23 is Phe, Pro, or Ser; Xaa at position 24 is Asn or Ala; Xaa at position 28 is Gly, Ala, Ser, Asp or Asn; Xaa at position 31 is Gln, Val, or Met; Xaa at position 32 is Asp or Ser; Xaa at position 35 is Met, Ile or Asp; Xaa at position 36 is Glu or Asp; Xaa at position 37 is Asn, Arg or Ser; Xaa at position 41 is Arg, Leu, or Thr; Xaa at position 42 is Pro or Ser; Xaa at position 45 is Glu or Leu; Xaa at position 46 is Ala or Ser; Xaa at position 48 is Asn, Val or Pro; Xaa at position 49 is Arg or His; Xaa at position 51 is Val or Ser; Xaa at position 53 is Ser, Asn, His or Gln; Xaa at position 55 is Gln or Glu; Xaa at position 59 is Ala or Gly; Xaa at position 62 is Ser, Ala or Pro; Xaa at position 65 is Lys, Arg or Ser; Xaa at position 67 is Leu, Glu, or Val; Xaa at position 68 is Leu, Glu, Val or Trp; Xaa at position 71 is Leu or Val; Xaa at position 73 is Leu, Ser or Tyr; Xaa at position 74 is Ala or Trp; Xaa at position 77 is Ala or Pro; Xaa at position 79 is Pro or Ser; Xaa at position 81 is His or Thr; Xaa at position 84 is His, Ile, or Thr; Xaa at position 86 is Lys or Arg; Xaa at position 87 is Asp, Ala or Met; Xaa at position 91 is Asn or Glu; Xaa at position 95 is Arg, Glu, Leu; Xaa at position 98 Thr or Gln; Xaa at position 102 is Lys, Val, Trp or Ser; Xaa at position 103 is Thr or Ser; Xaa at position 106 is Asn, Gln, or His; Xaa at position 109 is Ala or Glu; with the proviso that from four to forty-four of the amino acids designated by Xaa are different from the corresponding amino acids of native (15-125) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.
21 . A polypeptide according to claim 20 wherein Xaa at position 4 is Ile; Xaa at position 5 is Ala, or Ile; Xaa at position 6 is Pro, or Leu; Xaa at position 9 is Ala, or Leu; Xaa at position 11 is His; Xaa at position 15 is Arg or Val, or Ile; Xaa at position 18 is Ala or Asn, or Arg; Xaa at position 20 is Ser; Xaa at position 23 is Pro or Ser; Xaa at position 24 is Ala; Xaa at position 28 is Ala or Ser, or Asp, or Asn; Xaa at position 31 is Val or Met; and Xaa at position 32 is Ser.
22 . A polypeptide according to claim 20 wherein Xaa at position 35 is Ile, or Leu, or Asp; Xaa at position 36 is Asp; Xaa at position 37 is Arg, or Ser; Xaa at position 41 is Leu or Thr; Xaa at position 42 is Ser; Xaa at position 45 is Glu or Leu; Xaa at position 46 is Ala or Ser; Xaa at position 48 is Val, or Pro; Xaa at position 49 is His; Xaa at position 51 is Ser; Xaa at position 53 is Asn, or His, or Gln; and Xaa at position 55 is Glu.
23 . A polypeptide according to claim 20 wherein Xaa at position 59 is Gly; Xaa at position 62 is Ala, or Pro; Xaa at position 65 is Arg, or Ser; Xaa at position 67 is Gln or Val; Xaa at position 68 is Glu, or Val, or Trp; Xaa at position 71 is Val; Xaa at position 73 is Ser, or Tyr; Xaa at position 74 is Trp; Xaa at position 77 is Pro; Xaa at position 79 is Ser; Xaa at position 81 is Thr; Xaa at position 84 is Ile or Thr; Xaa at position 86 is Arg; Xaa at position 87 is Ala, or Met; and Xaa at position 91 is Glu.
24 . A polypeptide according to claim 20 wherein Xaa at position 95 is Glu, or Leu; Xaa at position 98 ia Gln; Xaa at position 102 is Val, or Trp, or Ser; Xaa at position 103 is Ser; Xaa at position 106 is Glu or His; and Xaa at position 109 is Glu.
25 . A polypeptide according to claim 20 wherein Xaa at position 4 is Ile; Xaa at position 5 is Ala, or Ile; Xaa at position 6 is Pro, or Leu; Xaa at position 9 is Ala, or Leu; Xaa at position 11 is His; Xaa at position 15 is Arg or Val, or Ile; Xaa at position 18 is Ala or Asn, or Arg; Xaa at position 20 is Ser; Xaa at position 23 is Pro or Ser; Xaa at position 24 is Ala; Xaa at position 28 is Ala or Ser, or Asp, or Asn; Xaa at position 31 is Val or Met; Xaa at position 32 is Ser; Xaa at position 35 is Ile, or Leu, or Asp; Xaa at position 36 is Asp; Xaa at position 37 is Arg, or Ser; Xaa at position 41 is Leu or Thr; Xaa at position 42 is Ser; Xaa at position 45 is Glu or Leu; Xaa at position 46 is Ala or Ser; Xaa at position 48 is Val, or Pro; Xaa at position 49 is His; Xaa at position 51 is Ser; Xaa at position 53 is Asn, or His, or Gln; and Xaa at position 55 is Glu.
26 . A polypeptide according to claim 20 wherein Xaa at position 59 is Gly; Xaa at position 62 is Ala, or Pro; Xaa at position 65 is Arg, or Ser; Xaa at position 67 is Gln or Val; Xaa at position 68 is Glu, or Val, or Trp; Xaa at position 71 is Val; Xaa at position 73 is Ser, or Tyr; Xaa at position 74 is Trp; Xaa at position 77 is Pro; Xaa at position 79 is Ser; Xaa at position 81 is Thr; Xaa at position 84 is Ile or Thr; Xaa at position 86 is Arg; Xaa at position 87 is Ala, or Met; Xaa at position 91 is Glu; Xaa at position 95 is Glu, or Lue; Xaa at position 98 is Gln; Xaa at position 102 is Val, or Trp, or Ser; Xaa at position 103 is Ser; Xaa at position 106 is Glu or His; and Xaa at position 109 is Glu.
27 . A polypeptide according to claim 20 which is selected from
Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:66]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Gln Asp Val Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn
Leu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser
Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu
Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly
Asp Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr
Leu Glu Asn Ala Gln Ala Gln Gln ;
Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys
[SEQ ID NO:67]
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Gln
Asp Met Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn Leu
Gln Ala Phe Asn Arg Ala Vai Lys Ser Leu Gln Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Gln Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys
[SEQ ID NO:68]
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Gln
Asp Met Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu
Alu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala
Ile Gb Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Glu Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:69]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu
Asp Gln Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu
Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Glu Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:70]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu
Asp Gln Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu
Glu Ser Phe Val Arq Ala Val Lys Asn Leu Glu Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Gln Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:71]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu
Asp Gln Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu
Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Glu Aso Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu Lys
[SEQ ID NO:72]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln
Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn Leu
Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly
Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala
Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp
Trp Gln Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Glu Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:73]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu
Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Aso Leu
Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly
Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala
Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp
Trp Gln Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Gln Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu Lys
[SEQ ID NO:74]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln
Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn Leu
Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala
Ile Gln Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu
Gln Gln Ala Gln Gln Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:75]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln
Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu
Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Ser Leu
Gln His Ala Gln Glu Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:76]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln
Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu
Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly
Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala
Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp
Trp Gln Glu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu
Gln Gln Ala Gln Gln Gln Gln;
Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu Lys
[SEQ ID NO:77]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln
Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu
Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly
Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala
Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp
Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu
Glu Gln Ala Gln Glu Gln Gln;
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
[SEQ ID NO:78]
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu
Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu
Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly
Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala
Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp
Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu
Glu His Ala Gln Glu Gln Gln;
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
[SEQ ID NO:79]
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu
Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu
Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Gln Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys
[SEQ ID NO:80]
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Gln
Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu
Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala
Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Gln Asn Ala Gln Ala Gln Gln;
Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys
[SEQ ID NO:81]
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Gln
Asp Met Asp Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn Leu
Leu Ala Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser Ala
Ile Gln Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp
Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu
Glu Asn Ala Gln Ala Gln Gln;
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu
[SEQ ID NO:82]
Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly
Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn
Leu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Glu Gln Gln;
Met Ala Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu
[SEQ ID NO:83]
Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly
Gln Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn
Leu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Gln Gln Gln;
Met Ala Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu
[SEQ ID NO:84]
Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly
Glu Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro Asn
Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Ser
Leu Glu His Ala Gln Gln Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO:85]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Gln Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Gln Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Ala Ile Gln Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu
Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly
Asp Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr
Leu Gln Asn Ala Gln Ala Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:86]
Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser
Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu
Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly
Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr
Leu Glu Asn Ala Gln Ala Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO:87]
Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu
Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly
Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr
Leu Glu Asn Ala Gln Ala Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO:88]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:89]
Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser
Gln Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Gln Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:90]
Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Glu Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:91]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Gln Asp Val Asp Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln;
Met Ala Asn Gys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:92]
Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Gln Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO:93]
Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser
Leu Glu His Ala Gln Glu Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO:94]
Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser
Leu Glu His Ala Gln Glu Gln Gln;
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn
[SEQ ID NO:95]
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Gln Gln Gln; and
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO:96]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Gln Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser
Leu Gln His Ala Gln Gln Gln Gln.
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO.: 296]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Gln Asp Val Asp Ile Leu Met Asp Arg Asn Leu Arg Leu Ser Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln.
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu
[SEQ ID NO.: 300]
Lys Arg Pro Pro Ala Pro Ser Leu Asp Pro Aso Asn Leu Asn Asp
Glu Asp Met Ser Ile Leu Met Gln ArgAsn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Gln Gln Gln
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO.: 301]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp
Gln Asp Met Ser Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn
Leu Gln Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Gln Gln Gln
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO.: 308]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala
Glu Asp Val Asp Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu
[SEQ ID NO.: 309]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp
Gln Asp Val Ser Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn
Leu Gln Ser Phe Val Arg Ala Val Lys Aso Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Aso Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Gln Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln
Met Ala Asa Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu
[SEQ ID NO.: 310]
Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp
Glu Asp Met Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn
Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asa Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr
Leu Glu Gln Ala Gln Gln Gln Gln
Met Ala Tyr Pro Gln Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn
[SEQ ID NO. 315]
Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys Arg Pro
Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Gln Asp Val
Asp Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Gln
Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala
Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln
Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln
Ala Gln Gln Gln Gln
Met Ala Tyr Pro Gln Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn
[SEQ ID NO.: 316]
Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys Arg Pro
Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Gln Asp Met
Asp Ile Leu Met Gln Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
Phe Val Arg Ala Val Lys His Leu Gln Asn Ala Ser Gly Ile Gln
Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala
Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln
Gln Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln Gln
Ala Gln Gln Gln Gln
Met Ala Asn Cys Ser Ile Met Ile Asp Gln Leu Ile His His Leu
[SEQ ID NO.: 318]
Lys Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser
Gln Asp Val Ser Ile Leu Met Gln Arg Asn Leu Arg Thr Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Gln Asn Ala Ser
Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly
Asp Trp Gln Glu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr
Leu Gln Gln Ala Gln Gln Gln Gln.
28 . A pharmaceutical composition for the treatment of hematopoietic cell deficiencies comprising a therapeutically effective amount of a mutant human interleukin-3 polypeptide selected from the group consisting of a polypeptide of claim 1 , a polypeptide of claim 2 , a polypeptide of claim 3 , a polypeptide of claim 4 , a polypeptide of claim 5 , a polypeptide of claim 6 , a polypeptide of claim 7 , a polypeptide of claim 8 , a polypeptide of claim 9 , a polypeptide of claim 10 , a polypeptide of claim 11 , a polypeptide of claim 12 , a polypeptide of claim 13 , a polypeptide of claim 14 , a polypeptide of claim 15 , a polypeptide of claim 16 , a polypeptide of claim 17; a polypeptide of claim 18 , a polypeptide of claim 19 , a polypeptide of claim 20 , a polypeptide of claim 21 , a polypeptide of claim 22 , a polypeptide of claim 23 , a polypeptide of claim 24 , a polypeptide of claim 25 , a polypeptide of claim 26 and a polypeptide of claim 27 , and a pharmaceutically acceptable carrier.
29 . A pharmaceutical composition according to claim 28 for the treatment of hematopoietic cell deficiencies comprising a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:88 and a pharmaceutically acceptable carrier.
30 . A pharmaceutical composition according to claim 28 for the treatment of hematopoietic cell deficiencies comprising a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:89 and a pharmaceutically acceptable carrier.
31 . A pharmaceutical composition according to claim 28 for the treatment of hematopoietic cell deficiencies comprising a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:90 and a pharmaceutically acceptable carrier.
32 . A pharmaceutical composition according to claim 28 for the treatment of hematopoietic cell deficiencies comprising a therapeutically effective amount of a polypeptide selected from the group consisting of
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:66;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:67;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:68;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:69;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:70;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:71;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:72;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:73;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:74;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:75;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:76;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:77;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:78;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:79;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:80;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:81;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:82;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:83;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:84;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:85;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:86;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:87;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:91;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:92;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:93;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:94;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:95;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:96;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:258;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:259;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:260;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:261;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:262;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:263;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:278;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:279;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:314;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:315;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:316;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:264;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:265;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:266;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:267;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:268;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:269;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:270;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:271;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:272;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:273;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:274;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:275;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:276;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:277;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:280;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:281;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:282;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:283;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:284;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:285;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:286;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:287;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:288;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:289;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:299;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:300;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:301;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:302;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:303;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:304;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:305;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:306;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:307;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:308;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:309;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:310;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:311;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:312;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:313;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:314;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:317;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:318;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:319;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:320;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:321;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:322;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:323;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:324;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:325;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:326;
and a pharmaceutically acceptable carrier.
33 . A method of stimulating the production of hematopoietic cells which comprises administering a therapeutically effective amount of a mutant human interleukin-3 polypeptide selected from the group consisting of a polypeptide of claim 1 , a polypeptide of claim 2 , a polypeptide of claim 3 , a polypeptide of claim 4 , a polypeptide of claim 5 , a polypeptide of claim 6 , a polypeptide of claim 7 , a polypeptide of claim 8 , a polypeptide of claim 9 , a polypeptide of claim 10 , a polypeptide of claim 11 , a polypeptide of claim 12 , a polypeptide of claim 13 , a polypeptide of claim 14 , a polypeptide of claim 15 , a polypeptide of claim 16 , a polypeptide of claim 17; a polypeptide of claim 18 , a polypeptide of claim 19 , a polypeptide of claim 20 , a polypeptide of claim 21 , a polypeptide of claim 22 , a polypeptide of claim 23 , a polypeptide of claim 24 , a polypeptide of claim 25 , a polypeptide of claim 26 , a polypeptide of claim 27 , to a patient in need of such treatment.
34 . A method according to claim 33 of stimulating the production of hematopoietic cells which comprises administering a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:88.
35 . A method according to claim 33 of stimulating the production of hematopoietic cells which comprises administering a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:89.
36 . A method according to claim 33 of stimulating the production of hematopoietic cells which comprises administering a therapeutically effective amount of a polypeptide having an amino acid sequence corresponding to SEQ ID NO:90.
37 . A method according to claim 33 of stimulating the production of hematopoietic cells which comprises administering a therapeutically effective amount of a polypeptide selected from the group consisting of
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:66;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:67;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:68;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:69;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:70;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:71;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:72;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:73;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:74;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:75;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:76;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:77;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:78;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:79;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:80;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:81;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:82;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:83;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:84;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:85;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:86;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:87;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:91;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:92;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:93;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:94;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:95;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:96;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:258;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:259;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:260;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:261;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:262;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:263;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:278;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:279;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:314;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:315;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:316;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:264;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:265;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:266;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:267;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:268;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:269;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:270;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:271;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:272;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:273;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:274;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:275;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:276;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:277;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:280;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:281;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:282;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:283;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:284;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:285;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:286;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:287;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:288;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:289;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:299;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:300;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:301;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:302;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:303;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:304;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:305;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:306;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:307;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:308;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:309;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:310;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:311;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:312;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:313;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:314;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:317;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:318;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:319;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:320;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:321;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:322;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:323;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:324;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:325;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:326;
to a patient in need of such treatment.
38 . A recombinant DNA sequence comprising vector DNA and a DNA that encodes a polypeptide selected from the group consisting of a polypeptide of claim 1 , a polypeptide of claim 2 , a polypeptide of claim 3 , a polypeptide of claim 4 , a polypeptide of claim 5 , a polypeptide of claim 6 , a polypeptide of claim 7 , a polypeptide of claim 8 , a polypeptide of claim 9 , a polypeptide of claim 10 , a polypeptide of claim 11 , a polypeptide of claim 12 , a polypeptide of claim 13 , a polypeptide of claim 14 , a polypeptide of claim 15 , a polypeptide of claim 16 , a polypeptide of claim 17; a polypeptide of claim 18 , a polypeptide of claim 19 , a polypeptide of claim 20 , a polypeptide of claim 21 , a polypeptide of claim 22 , a polypeptide of claim 23 , a polypeptide of claim 24 , a polypeptide of claim 25 , a polypeptide of claim 26 , or a polypeptide of claim 27 .
39 . A recombinant DNA sequence according to claim 38 comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:97.
40 . A recombinant DNA sequence according to claim 38 comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:100 or 103.
41 . A recombinant DNA sequence according to claim 38 comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:161.
42 . A recombinant DNA sequence according to claim 38 comprising vector DNA and a DNA selected from
a DNA having a nucleotide sequence corresponding to SEQ ID NO:98;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:99;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:101;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:102;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:104;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:105;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:106;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:107;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:108;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:109;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:110;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:111;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:112;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:113;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:114;
a DNA having a nucleotide sequence corresponding to SEQ ID NO: 115;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:116;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:117;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:118;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:119;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:120;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:121;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:122;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:123;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:124;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:125;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:126;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:127;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:160;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:161;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:398;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:399;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:346;
a DNA having a nucleotide sequence corresponding to SEQ ID NO:347
a DNA having a nucleotide sequence corresponding to SEQ ID NO:303
a DNA having a nucleotide sequence corresponding to SEQ ID NO:404
a DNA having a nucleotide sequence corresponding to SEQ ID NO:405
a DNA having a nucleotide sequence corresponding to SEQ ID NO:332
a DNA having a nucleotide sequence corresponding to SEQ ID NO:333
a DNA having a nucleotide sequence corresponding to SEQ ID NO:334
a DNA having a nucleotide sequence corresponding to SEQ ID NO:335
a DNA having a nucleotide sequence corresponding to SEQ ID NO:336
a DNA having a nucleotide sequence corresponding to SEQ ID NO:337
a DNA having a nucleotide sequence corresponding to SEQ ID NO:338
a DNA having a nucleotide sequence corresponding to SEQ ID NO:339
a DNA having a nucleotide sequence corresponding to SEQ ID NO:340
a DNA having a nucleotide sequence corresponding to SEQ ID NO:341
a DNA having a nucleotide sequence corresponding to SEQ ID NO:342
a DNA having a nucleotide sequence corresponding to SEQ ID NO:343
a DNA having a nucleotide sequence corresponding to SEQ ID NO:344
a DNA having a nucleotide sequence corresponding to SEQ ID NO:345
a DNA having a nucleotide sequence corresponding to SEQ ID NO:348
a DNA having a nucleotide sequence corresponding to SEQ ID NO:349
a DNA having a nucleotide sequence corresponding to SEQ ID NO:350
a DNA having a nucleotide sequence corresponding to SEQ ID NO:352
a DNA having a nucleotide sequence corresponding to SEQ ID NO:353
a DNA having a nucleotide sequence corresponding to SEQ ID NO:354
a DNA having a nucleotide sequence corresponding to SEQ ID NO:355
a DNA having a nucleotide sequence corresponding to SEQ ID NO:356
a DNA having a nucleotide sequence corresponding to SEQ ID NO:357
a DNA having a nucleotide sequence corresponding to SEQ ID NO:358
a DNA having a nucleotide sequence corresponding to SEQ ID NO:359
a DNA having a nucleotide sequence corresponding to SEQ ID NO:360
a DNA having a nucleotide sequence corresponding to SEQ ID NO:361
a DNA having a nucleotide sequence corresponding to SEQ ID NO:362
a DNA having a nucleotide sequence corresponding to SEQ ID NO:363
a DNA having a nucleotide sequence corresponding to SEQ ID NO:364
a DNA having a nucleotide sequence corresponding to SEQ ID NO:365
a DNA having a nucleotide sequence corresponding to SEQ ID NO:366
a DNA having a nucleotide sequence corresponding to SEQ ID NO:367
a DNA having a nucleotide sequence corresponding to SEQ ID NO:368
a DNA having a nucleotide sequence corresponding to SEQ ID NO:369
a DNA having a nucleotide sequence corresponding to SEQ ID NO:370
a DNA having a nucleotide sequence corresponding to SEQ ID NO:371
a DNA having a nucleotide sequence corresponding to SEQ ID NO:372
a DNA having a nucleotide sequence corresponding to SEQ ID NO:373
a DNA having a nucleotide sequence corresponding to SEQ ID NO:374
a DNA having a nucleotide sequence corresponding to SEQ ID NO:375
a DNA having a nucleotide sequence corresponding to SEQ ID NO:376
a DNA having a nucleotide sequence corresponding to SEQ ID NO:377
a DNA having a nucleotide sequence corresponding to SEQ ID NO:378
a DNA having a nucleotide sequence corresponding to SEQ ID NO:379
a DNA having a nucleotide sequence corresponding to SEQ ID NO:380
a DNA having a nucleotide sequence corresponding to SEQ ID NO:381
a DNA having a nucleotide sequence corresponding to SEQ ID NO:382
a DNA having a nucleotide sequence corresponding to SEQ ID NO:384
a DNA having a nucleotide sequence corresponding to SEQ ID NO:385
a DNA having a nucleotide sequence corresponding to SEQ ID NO:386
a DNA having a nucleotide sequence corresponding to SEQ ID NO:387
a DNA having a nucleotide sequence corresponding to SEQ ID NO:388
a DNA having a nucleotide sequence corresponding to SEQ ID NO:389
a DNA having a nucleotide sequence corresponding to SEQ ID NO:390
a DNA having a nucleotide sequence corresponding to SEQ ID NO:391
a DNA having a nucleotide sequence corresponding to SEQ ID NO:392
43 . A host cell containing a recombinant DNA sequence of claim 38 and capable of expressing the encoded polypeptide.
44 . A host cell of claim 43 containing a recombinant DNA vector comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:97 and capable of expressing the encoded polypeptide.
45 . A host cell of claim 43 containing a recombinant DNA vector comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:100 or 103 and capable of expressing the encoded polypeptide.
46 . A host cell of claim 43 containing a recombinant DNA vector comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:161 and capable of expressing the encoded polypeptide.
47 . A method of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA sequence of claim 38 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and (b) harvesting the polypeptide from the culture.
48 . A method according to claim 47 of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:97 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and
(b) harvesting the polypeptide from the culture.
49 . A method according to claim 47 of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:100 or 103 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and
(b) harvesting the polypeptide from the culture.
50 . A method according to claim 47 of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:161 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and
(b) harvesting the polypeptide from the culture.
51 . A vector containing a gene having a DNA sequence selected from the group consisting of:
a DNA having a nucleotide sequence corresponding to SEQ ID NO:97; a DNA having a nucleotide sequence corresponding to SEQ ID NO:100; a DNA having a nucleotide sequence corresponding to SEQ ID NO:103; a DNA having a nucleotide sequence corresponding to SEQ ID NO:160; a DNA having a nucleotide sequence corresponding to SEQ ID NO:161; a DNA having a nucleotide sequence corresponding to SEQ ID NO:404; a DNA having a nucleotide sequence corresponding to SEQ ID NO:405; a DNA having a nucleotide sequence corresponding to SEQ ID NO:364; a DNA having a nucleotide sequence corresponding to SEQ ID NO:368; a DNA having a nucleotide sequence corresponding to SEQ ID NO:369; a DNA having a nucleotide sequence corresponding to SEQ ID NO:376; a DNA having a nucleotide sequence corresponding to SEQ ID NO:377; a DNA having a nucleotide sequence corresponding to SEQ ID NO:378; a DNA having a nucleotide sequence corresponding to SEQ ID NO:385;
52 . A recombinant DNA vector comprising a promoter, a ribosome binding site, and a signal peptide directly linked to a DNA sequence encoding a polypeptide selected from the group consisting of
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:88; a polypeptide having an amino acid sequence corresponding to SEQ ID NO:89; and a polypeptide having an amino acid sequence corresponding to SEQ ID NO:90; said vector being capable of directing expression of said mutant human interleukin-3 polypeptide.
53 . A recombinant DNA vector according to claim 51 wherein the promoter is AraBAD.
54 . A recombinant DNA vector according to claim 51 wherein the ribosome binding site is g10-L.
55 . A recombinant DNA vector according to claim 51 wherein the signal peptide is a lamB signal peptide.
56 . A recombinant DNA vector according to claim 51 wherein the signal peptide is the lamB signal peptide depicted in FIG. 8.
57 . A recombinant DNA vector according to claim 51 wherein the promoter is AraBAD and the ribosome binding site is g10-L.
58 . A recombinant DNA vector according to claim 51 wherein the promoter is AraBAD, the ribosome binding site is g10-L, and the signal peptide is a lamB signal peptide.
59 . A recombinant DNA vector according to claim 51 wherein the promoter is AraBAD, the ribosome binding site is g10-L, and the signal peptide is the lamB signal peptide depicted in FIG. 8.
60 . A recombinant bacterial host which comprises the vector of claim 51 wherein said host secretes a mutant human interleukin-3 polypeptide selected from the group consisting of
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:88;
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:89; and
a polypeptide having an amino acid sequence corresponding to SEQ ID NO:90.
61 . A polypeptide of the formula
1 5 10
[SEQ ID NO:129]
(Met) m -Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr
15 20
Ser Trp Val Asn Cys Ser Xaa Met Ile Asp Glu Ile Ile
25 30 35
Xaa His Leu Lys Xaa Pro Pro Xaa Pro Leu Leu Asp Xaa
40 45 50
Asn Asn Leu Asn Xaa Glu Asp Xaa Asp Ile Leu Met Glu
55 60
Xaa Asn Leu Arg Xaa Pro Asn Leu Xaa Xaa Phe Xaa Arg
65 70 75
Ala Val Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa
80 85
Ile Leu Xaa Asn Leu Xaa Pro Cys Leu Pro Xaa Ala Thr
90 95 100
Ala Ala Pro Xaa Arg His Pro Ile Xaa Ile Lys Xaa Gly
105 110 115
Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Thr Phe Tyr Leu
120 125
Xaa Thr Leu Glu Xaa Ala Gln Xaa Gln Gln Thr Thr Leu
130
Ser Leu Ala Ile Phe
wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa at position 25 is Thr or His; Xaa at position 29 is Gln, Arg, or Val; Xaa at position 32 is Leu, Ala, or Asn; Xaa at position 37 is Phe, Pro, or Ser; Xaa at position 42 is Glu, Ala, or Ser; Xaa at position 45 is Gln, Val, or Met; Xaa at position 51 is Asn or Arg; Xaa at position 55 is Arg, Leu, or Thr; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa at position 62 is Asn or Val; Xaa at position 67 is Ser, Asn, or His; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Ala or Gly; Xaa at position 76 is Ser or Ala; Xaa at position 79 is Lys or Arg; Xaa at position 82 is Leu, Glu, or Val; Xaa at position 87 is Leu or Ser; Xaa at position 93 is Pro or Ser; Xaa at position 98 is His, Ile, or Thr; Xaa at position 101 is Asp or Ala; Xaa at position 105 is Asn or Glu; Xaa at position 109 is Arg or Glu; Xaa at position 116 is Lys or Val; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123 is Ala or Glu; with the proviso that from four to twenty-seven of the amino acids designated by Xaa are different from the corresponding amino acids of native human interleukin-3 and wherein from 1 to 14 of amino acids 1 to 14 has been deleted from the N-terminus and/or from 1 to 15 of amino acids 119 to 133 has been deleted from the C-terminus of the polypeptide; or a polypeptide having substantially the same structure and substantially the same biological activity.
62 . A method according to claim 47 of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:160 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and
(b) harvesting the polypeptide from the culture.
63 . A method according to claim 47 of producing a mutant human interleukin-3 polypeptide comprising the steps of:
(a) culturing a host cell containing a recombinant DNA sequence comprising vector DNA and a DNA having a nucleotide sequence corresponding to SEQ ID NO:161 and capable of expressing the encoded polypeptide under conditions permitting expression of the recombinant DNA; and
(b) harvesting the polypeptide from the culture.
64 . A host cell containing a recombinant DNA vector comprising vector DNA and a DNA sequence selected from the group consisting of:
a DNA having a nucleotide sequence corresponding to SEQ ID NO:160; and a DNA having a nucleotide sequence corresponding to SEQ ID NO:161; and capable of expressing the encoded polypeptide.
65 . A polypeptide according to claim 27 which is:
[SEQ ID NO:89]
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile
His His Leu
Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn
Leu Asn Ser
Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg
Thr Pro Asn
Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu
Asn Ala Ser
Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys
Leu Pro Ser
Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile
Lys Ala Gly
Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr
Leu Val Thr
Leu Glu Gln Ala Gln Glu Gln Gln.
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