US2003054426A1PendingUtilityA1
Peptide biomarker and method of identification
Priority: Aug 31, 2001Filed: Aug 30, 2002Published: Mar 20, 2003
Est. expiryAug 31, 2021(expired)· nominal 20-yr term from priority
G01N 2800/102C12Q 1/37Y10T436/105831G01N 2800/52G01N 2800/105C07K 16/18C07K 14/78G01N 33/6821G01N 2500/02G01N 2333/78G01N 33/6887G01N 33/6842G01N 2333/96486
32
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Claims
Abstract
A method for identifying and quantifying peptides resulting from enzyme cleavage of collagen type II by mass spectrometric analysis to detect characteristic peptide fragments of a carboxy-terminus having known mass to charge ratios to identify the peptide having that carboxy-terminus. Identification and quantification of the peptides in a biological sample is used to assess activity of proteolytic enzymes in diseases or physiological conditions such as osteoarthritis and rheumatoid arthritis and to assess the efficacy of enzyme blocking agents.
Claims
exact text as granted — not AI-modified1 . A method of detecting enzymatic proteolysis of a collagen, the method comprising identifiying and quantifying a specific peptide proteolysis product of the collagen occurring in a biological sample or biological extract.
2 . The method of claim 1 wherein the collagen is type II collagen and the specific peptide has a C-terminus represented by Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 1) and post-translational modifications thereof.
3 . The method of claim 1 wherein the collagen is type I collagen and the specific peptide has a C-terminus represented by Gly-Thr-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 12) and post-translational modifications thereof.
4 . The method of claim 1 wherein the collagen is type III collagen and the specific peptide has a C-terminus represented by Gly-Ala-Pro-Gly-Pro-Leu-Gly (SEQ ID NO: 13) and post-translational modifications thereof.
5 . The method of claim 1 wherein the enzymatic proteolysis of a collagen is the result of collagenase activity.
6 . The method of claim 5 wherein the collagenase is matrix metalloproteinase-13.
7 . The method of claim 5 wherein the method comprises monitoring changes in collagenase activity caused by a drug.
8 . The method of claim 1 comprising identifying and quantifying more than one specific peptide occurring in the biological sample or biological extract.
9 . The method of claim 1 wherein the proteolysis product of the collagen occurs in a biological sample selected from the group consisting of synovial fluid, whole blood, plasma and urine.
10 . The method of claim 1 wherein the specific peptide proteolysis product is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
11 . The method of claim 10 wherein the biological specimen or biological extract is from a human subject, the method comprising the identification and quantification of the peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 2) or its post-translationally-modified analogs in a biological sample or biological extract.
12 . The method of claim 10 wherein the biological specimen or biological extract is from a human subject, the method comprising the identification and quantification of the peptide Leu-Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 15) or post-translational modifications thereof in a biological sample or biological extract.
13 . The method of claim 10 wherein the biological specimen or biological extract is from a human subject, the method comprising the identification and quantification of the peptide A la-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 16) or post-translational modifications thereof in a biological sample or biological extract.
14 . The method of claim 10 wherein the biological specimen or biological extract is from a human subject, the method comprising the identification and quantification of the peptide Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 17) or post-translational modifications thereof in a biological sample or biological extract.
15 . The method of claim 10 wherein the biological specimen or biological extract is from a bovine subject, the method comprising the identification and quantification of a peptide in a biological sample or biological extract, the peptide being selected from the group consisting of Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 3) and post-translational modifications thereof, Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 34) and post-translational modifications thereof, Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 35) and post-translational modifications thereof, Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 36) and post-translational modifications thererof, Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 37) and post-translational modifications thereof, and Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 38) and post-translational modifications thereof.
16 . The method of claim 10 wherein the biological specimen or biological extract is from a canine subject, the method comprising the identification and quantification of a peptide in a biological sample or biological extract, the peptide being selected from the group consisting of Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 4) and post-translational modifications thereof, Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 18) and post-translational modifications thereof, Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 19) and post-translational modifications thereof, Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 20) and post-translational modifications thereof, Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 21) and post-translational modifications thereof, and Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 22) and post-translational modifications thereof.
17 . The method of claim 10 wherein the biological specimen or biological extract is from an equine subject, the method comprising the identification and quantification of a peptide in a biological sample or biological extract, the peptide being selected from the group consisting of Leu-Gln-Gly-Pro-Ala-Gly-PrO-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 11) and post-translational modifications thereof, Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 29) and post-translational modifications thereof, Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 30) and post-translational modifications thereof, Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 31) and post-translational modifications thereof, Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 32) and post-translational modifications thereof, and Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 33) and post-translational modifications thereof.
18 . The method of claim 10 wherein the biological specimen or biological extract is from a rat, the method comprising the identification and quantification of a peptide in a biological sample or biological extract, the peptide being selected from the group consisting of Leu-Gln-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 8) and post-translational modifications thereof, Ala-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 39) and post-translational modifications thereof, Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 40) and post-translational modifications thereof, Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 41) and post-translational modifications thereof, and Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 42) and post-translational modifications thereof.
19 . The method of claim 10 wherein the biological specimen or biological extract is from a feline subject, the method comprising the identification and quantification of a peptide in a biological sample or biological extract, the peptide being selected from the group consisting of Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 23) and post-translational modifications thereof, Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 24) and post-translational modifications thereof, Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 25) and post-translational modifications thereof, Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 26) and post-translational modifications thereof, Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 27) and post-translational modifications thereof, and Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 28) and post-translational modifications thereof.
20 . The method of claim 10 wherein the biological specimen or biological extract is from a guinea pig, the method comprising the identification and quantification of the peptide Val-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Val-Ser-Gly-Ala-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 43) or post-translational modifications thereof in a biological sample or biological extract.
21 . The method of claim 10 further comprising forming a derivative or modification of the specific peptide proteolysis product in the sample or extract before identifying and quantifying the peptide.
22 . The method of claim 1 further comprising forming a derivative or modification of the specific peptide proteolysis product in the sample or extract before identifying and quantifying the peptide.
23 . A method of identifying the amino acid sequence of a collagen degradation peptide in a biological sample comprising:
determining a relative molecular mass of the peptide; fragmenting the peptide by collisional activation to create characteristic diagnostic sequences of a C-terminus of the peptide; identifying the C-terminus of the peptide by identification of the characteristic diagnostic sequences, said diagnostic sequences having known mass to charge ratios; and identifying an N-terminal sequence of the peptide by determining an amino acid sequence that when combined with the identified C-terminal sequence has the relative molecular mass of the peptide, thereby identifying the collagen degradation peptide.
24 . The method of claim 23 wherein the C-terminus is Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 1) and post-translational modifications thereof.
25 . The method of claim 23 wherein the C-terminus is Gly-Thr-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 12) and post-translational modifications thereof.
26 . The method of claim 23 wherein the C-terminus is Gly-Ala-Pro-Gly-Pro-Leu-Gly (SEQ ID NO: 13) and post-translational modifications thereof.
27 . The method of claim 23 wherein the step of identifying an N-terminal sequence of the peptide by determining an amino sequence that when combined with the identified C-terminal sequence has the relative molecular mass of the peptide further comprises determining an amino acid sequence that when combined with the identified C-terminal sequence has the mass of a peptide in a data base comprising known peptides.
28 . The method of claim 23 wherein the step of identifying a C-terminus of the peptide by identification of characteristic diagnostic sequences further comprises identifying three diagnostic sequences having the sequences Pro-Gln-Gly (SEQ ID NO: 5), Pro-Gly-Pro-Gln-Gly (SEQ.ID NO: 6) and Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 7) and post-translational modifications of the sequences.
29 . The method of claim 28 wherein the three diagnostic sequences have a mass to charge ratio of approximately 301, approximately 471 and approximately 568, respectively.
30 . The method of claim 28 wherein the three diagnostic sequences have a mass to charge ratio of approximately 301, approximately 455 and approximately 552, respectively.
31 . A method of identifying the amino acid sequence of a collagen degradation peptide having an N-terminal segment and a carboxy-terminus (C-terminus) from a protein of known sequence comprising the steps of:
determining a relative molecular mass of the peptide; fragmenting the peptide to produce fragment ions; identifying known fragment ions from the fragmentation of the C-terminus of the peptide; identifying an amino acid sequence of the C-terminus from the identified ion fragments from the C-terminus of the peptide; determining an amino acid sequence of an N-terminus of the peptide from the amino acid sequence of the C-terminus and the relative molecular mass of the peptide; determining an amino acid sequence of the peptide from the amino acid sequence of the N-terminus and the amino acid sequence of the C-terminus.
32 . The method of claim 31 wherein the collagen degradation peptide comprises a proteolytic enzyme cleavage product.
33 . The method of claim 31 comprising a further step of determining at least one site and type of post-translational modification to the peptide from the identified ion fragments from the peptide.
34 . The method of claim 31 wherein the proteolytic enzyme is a collagenase or matrix metalloproteinase.
35 . The method of claim 31 wherein the collagen degradation peptide is a collagen type I degradation peptide.
36 . The method of claim 31 wherein the collagen degradation peptide is a collagen type III degradation peptide.
37 . The method of claim 22 wherein the matrix metalloproteinase is matrix metalloproteinase-13.
38 . The method of claim 17 wherein the steps of determining a relative molecular mass of the peptide, fragmenting the peptide and identifying known peptide ion fragments from the C-terminus of the peptide are performed by tandem mass spectrometry.
39 . An isolated peptide selected from the group consisting of the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 2) and post-translationally modified analogs thereof, the isolated peptide Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 15) and post-translationally modified analogs thereof, the isolated peptide A la-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 16) and post-translationally modified analogs thereof, the isolated peptide Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 17) and post-translationally modified analogs thereof, the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 3) and post-translationally modified analogs thereof, the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 8) and post-translationally modified analogs thereof, the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 11) and post-translationally modified analogs thereof, the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 3) and post-translationally modified analogs thereof, the isolated peptide Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 34) and post-translationally modified analogs thereof, the isolated peptide Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 35) and post-translationally modified analogs thereof, the isolated peptide Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 36) and post-translationally modified analogs thereof, the isolated peptide Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 37) and post-translationally modified analogs thereof, the isolated peptide Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 38) and post-translationally modified analogs thereof, the isolated peptide Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 18) and post-translationally modified analogs thereof, the isolated peptide Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 19) and post-translationally modified analogs thereof, the isolated peptide Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 20) and post-translationally modified analogs thereof, the isolated peptide Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 21) and post-translationally modified analogs thereof, the isolated peptide Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 22) and post-translationally modified analogs thereof, the isolated peptide Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 23) and post-translationally modified analogs thereof, the isolated peptide Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 24) and post-translationally modified analogs thereof, the isolated peptide Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 25) and post-translationally modified analogs thereof, the isolated peptide Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 26) and post-translationally modified analogs thereof, the isolated peptide Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 27) and post-translationally modified analogs thereof, the isolated peptide Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 28) and post-translationally modified analogs thereof, the isolated peptide Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 29) and post-translationally modified analogs thereof, the isolated peptide Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 30) and post-translationally modified analogs thereof, the isolated peptide Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 31) and post-translationally modified analogs thereof, the isolated peptide Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 32) and post-translationally modified analogs thereof, the isolated peptide Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 33) and post-translationally modified analogs thereof, the isolated peptide Ala-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 39) and post-translationally modified analogs thereof, the isolated peptide Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 40) and post-translationally modified analogs thereof, the isolated peptide Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 41) and post-translationally modified analogs thereof, the isolated peptide Val-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Val-Ser-Gly-Ala-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 43) and post-translationally modified analogs thereof, the isolated peptide Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 42) and post-translationally modified analogs thereof.
40 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 3) or a post-translationally modified analog thereof.
41 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 8) or a post-translationally modified analog thereof.
42 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 11) or a post-translationally modified analog thereof.
43 . The artificial peptide Val-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 14) or a post-translationally modified analog thereof.
44 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 3) or a post-translationally modified analog thereof.
45 . The isolated peptide of claim 39 wherein the peptide is Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 34) or a post-translationally modified analog thereof.
46 . The isolated peptide of claim 39 wherein the peptide is Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 35) or a post-translationally modified analog thereof.
47 . The isolated peptide of claim 39 wherein the peptide is Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 36) or a post-translationally modified analog thereof.
48 . The isolated peptide of claim 39 wherein the peptide is Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 37) or a post-translationally modified analog thereof.
49 . The isolated peptide of claim 39 wherein the peptide is Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 38) or a post-translationally modified analog thereof.
50 . The isolated peptide of claim 39 wherein the peptide is Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 18) or a post-translationally modified analog thereof.
51 . The isolated peptide of claim 39 wherein the peptide is Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 19) or a post-translationally modified analog thereof.
52 . The isolated peptide of claim 39 wherein the peptide is Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 20) or a post-translationally modified analog thereof.
53 . The isolated peptide of claim 39 wherein the peptide is Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 21) or a post-translationally modified analog thereof.
54 . The isolated peptide of claim 39 wherein the peptide is Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 22) or a post-translationally modified analog thereof.
55 . The isolated peptide of claim 39 wherein the peptide is Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 23) or a post-translationally modified analog thereof.
56 . The isolated peptide of claim 39 wherein the peptide is Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 24) or a post-translationally modified analog thereof.
57 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 25) or a post-translationally modified analog thereof.
58 . The isolated peptide of claim 39 wherein the peptide is Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 26) or a post-translationally modified analog thereof.
59 . The isolated peptide of claim 39 wherein the peptide is Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 27) or a post-translationally modified analog thereof.
60 . The isolated peptide of claim 39 wherein the peptide is Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 28) or a post-translationally modified analog thereof.
61 . The isolated peptide of claim 39 wherein the peptide is Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 29) or a post-translationally modified analog thereof.
62 . The isolated peptide of claim 39 wherein the peptide is Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 30) or a post-translationally modified analog thereof.
63 . The isolated peptide of claim 39 wherein the peptide is Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 31) or a post-translationally modified analog thereof.
64 . The isolated peptide of claim 39 wherein the peptide is Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 32) or a post-translationally modified analog thereof.
65 . The isolated peptide of claim 39 wherein the peptide is Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Pro-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 33) or a post-translationally modified analog thereof.
66 . The isolated peptide of claim 39 wherein the peptide is Ala-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 39) or a post-translationally modified analog thereof.
67 . The isolated peptide of claim 39 wherein the peptide is Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 40) or a post-translationally modified analog thereof.
68 . The isolated peptide of claim 39 wherein the peptide is Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 41) or a post-translationally modified analog thereof.
69 . The isolated peptide of claim 39 wherein the peptide is Val-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Asp-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Val-Ser-Gly-Ala-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 43) or a post-translationally modified analog thereof.
70 . The isolated peptide of claim 39 wherein the peptide is Asp-Gly-Pro-Ser-Gly-Ser-Asp-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 42) or a post-translationally modified analog thereof.
71 . The isolated peptide of claim 39 wherein the peptide is Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 2) or a post-translationally modified analog thereof.
72 . The isolated peptide of claim 39 wherein the peptide is Lys-Gly-Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 15) or a post-translationally modified analog thereof.
73 . The isolated peptide of claim 39 wherein the peptide is Ala-Arg-Gly-Asp-Ser-Gly-Pro-Pro-Gly-Arg-Ala-Gly-Glu-Pro-Gly-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 16) or a post-translationally modified analog thereof.
74 . The isolated peptide of claim 39 wherein the peptide is Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 17) or a post-translationally modified analog thereof.
75 . The artificial peptide Val-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 14) or a post-translationally modified analog thereof.
76 . A biomarker of collagenase enzyme activity comprising a peptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
77 . The biomarker of claim 71 wherein the peptide has been derivativized or modified to form an identifiable derivative or modification thereof for quantifying the biomarker.
78 . An antibody directed against an N-terminus sequence of at least seven amino acids of a peptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
79 . A biomarker of collagenase enzyme inhibition comprising a peptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 1, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
80 . The biomarker of claim 79 wherein the peptide has been derivativized or modified to form an identifiable derivative or modification thereof for quantifying the biomarker.
81 . A method of determining the efficacy of a drug or agent used to reduce the proteolytic activity of a collagenase enzyme comprising the identification and quantification of a collagen II degradation fragment in a biological fluid sample having a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
82 . The method of claim 81 , further comprising forming a derivative or modification of the collagen II degradation fragment in the sample or extract before identifying and quantifying the degradation fragment.
83 . A method of diagnosing a disease or condition in a subject characterized by collagen type II degradation comprising identifying and quantifying a peptide in a biological sample, the peptide being selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 and post-translationally modified analogs of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43.
84 . The method of claim 83 further comprising forming a derivative or modification of the peptide in the sample or extract before identifying and quantifying the peptide.
85 . The method of claim 83 wherein the disease or condition characterized by collagen type II degradation is selected from the group consisting of osteoarthritis and rheumatoid arthritis.
86 . The method of claim 83 wherein the biological sample is selected from the group consisting of urine, blood, plasma and synovial fluid.
87 . A method of determining the efficacy of a drug or agent in a human or animal subject used to reduce or eliminate the proteolytic activity of a collagenase enzyme in a disease or condition characterized by collagen type II degradation comprising the identification and quantification of a collagen type II degradation peptide in a biological sample from the subject, the collagen type II degradation peptide being selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43, and post-translational analogs of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43.
88 . The method of claim 87 wherein the collagenase enzyme is a matrix metalloproteinase.
89 . The method of claim 88 wherein the matrix metalloproteinase is selected from the group consisting of matrix metalloproteinase-1, matrix metalloproteinase-8 and matrix metalloproteinase-13.
90 . The method of claim 88 wherein the drug or agent used to reduce the proteolytic activity of a collagenase enzyme is a matrix metalloproteinase inhibitor.
91 . A method for determining cartilage damage in a physiological condition in a mammal characterized in part by cartilage damage comprising analyzing a biological sample of the mammal to determine the quantity of a marker peptide having sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, and post-translational analogs of SEQ ID NO: 2, of SEQ ID NO: 3 and SEQ ID NO: 4, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43.
92 . The method of claim 90 wherein the identification and quantification of the marker peptide further comprises the following steps:
determining a mass of the marker peptide;
fragmenting the marker peptide by collisional activation;
identifying a C-terminal sequence Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID. NO: 1) and a post-translationally modified analog thereof by identification of fragments of the marker peptide, said fragments identified by known mass to charge ratios;
identifying an N-terminal sequence of a peptide that when combined with the a C-terminal sequence Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID. NO: 1) and the post-translationally modified analogs thereof has the determined mass of the marker peptide, thereby identifying the marker peptide; and
determining a relative amount of the marker peptide in the body fluid.
93 . The method of claim 92 wherein the biological sample is selected from the group of biological fluids consisting of urine, blood, plasma and synovial fluid.
94 . The method of claim 92 wherein the fragments with known mass to charge ratios are fragment SEQ ID NO: 5 having a mass to charge ratio of approximately 301, the fragment sequence SEQ ID NO: 6 having a mass to charge ratio of approximately 455, and the fragment SEQ ID NO: 7 having a mass to charge ratio of approximately 552.
95 . The method of claim 91 wherein the fragments with known mass to charge ratios are fragment SEQ ID NO: 5 having a mass to charge ratio of approximately 301, the fragment sequence SEQ ID NO: 6 having a mass to charge ratio of approximately 471, and the fragment SEQ ID NO: 7 having a mass to charge ratio of approximately 568.
96 . The method of claim 91 wherein the physiological condition in a mammal characterized in part by cartilage damage is osteoarthritis.
97 . The method of claim 91 wherein the physiological condition in a mammal characterized in part by cartilage damage is rheumatoid arthritis.
98 . The method of claim 97 wherein the steps of determining the relative amount of the marker peptide in the biological sample further comprises:
adding known quantities of a standard peptide to the biological sample;
plotting an abundance of the standard peptide in the biological sample versus an analytical response to obtain a graphic curve of the abundance/response of the standard peptide;
plotting an abundance of the marker peptide in the biological sample to obtain a graphic curve of the abundance of the marker peptide;
determining an area under the graphic curve of the abundance of the marker peptide; and
comparing the area under the graphic curve of the abundance of the marker peptide to the area under the graphic curve of the abundance of the standard peptide to obtain a relative of abundance of the marker peptide in the biological sample.
99 . The method of claim 98 further comprising comparing the analytical response of the marker peptide to an internal standard peptide to determine a predetermined quantity of marker peptide in the biological sample.
100 . The method of claim 98 wherein the internal standard peptide is an isotopically-labelled homolog of the marker peptide.
101 . The method of claim 92 wherein the internal standard peptide is a peptide having a similar structure to, or biological properties as, the marker peptide.
102 . A method for determining collagen degradation in a physiological condition in a mammal characterized in part by collagen degradation comprising analyzing a biological sample of the mammal to determine a quantity of peptide comprising fragment SEQ ID NO: 13 and post-translational analogs of SEQ ID NO: 13.
103 . A method for determining collagen degradation in a physiological condition in a mammal characterized in part by collagen degradation comprising analyzing a biological sample of the mammal to determine a quantity of peptide comprising fragment SEQ ID NO: 12 and post-translational analogs of SEQ ID NO: 12.
104 . A method of identifying and quantifying target collagen degradation peptides in a biological sample, comprising:
obtaining a biological sample; spiking the biological sample with internal standard; spiking unknown samples with internal standard; centrifuging each sample; retaining solute solution from each sample adding formic acid to the retentate; centrifuging until a predetermine volume of the solute solution remains; transferring the solute solution to a reaction-vial; evaporating the solute solution to dryness under nitrogen; dissolving each dryed sample in water containing formic acid; transferring each dried sample to an autosampler vial; injecting a sample aliquot from the autosampler vial onto a column connected to a MS/MS interfaced spectrometer; eluting the sample aliquot; holding for approximately 2 minutes; re-equilibrating the column; monitoring ion pairs for the target peptides and internal standard peptides; integrating peak areas of the ion pairs for each target peptide; normalizing the integrated peak areas of the target peptides; and comparing the normalized integrated peaks for the target peptides to a standard calibration curve to determine the quantities of the target peptides in each unknown sample.
105 . The method of claim 104 wherein the target collagen degradation peptides are selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO. 8, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, and post-translational analogs of SEQ ID NO: 2, of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO. 8, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43.
106 . The method of claim 104 wherein the biological sample is urine.
107 . The peptide Gly-Ala-Pro-Gly-Pro-Leu-Gly (SEQ ID NO: 13), or a post-translationally-modified analog thereof.
108 . The peptide Val-Leu-Gln-Gly-Pro-Ala-Gly-Pro-Pro-Gly-Glu-Lys-Gly-Glu-Pro-Gly-Asp-Asp-Gly-Pro-Ser-Gly-Ala-Glu-Gly-Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 14) or a post-translationally modified analog thereof.
109 . A method of detecting the enzymatic degradation of collagen type II from a subject, the method comprising the identification and quantification of identifying three diagnostic sequences having the sequences Pro-Gln-Gly (SEQ ID NO: 5), Pro-Gly-Pro-Gln-Gly (SEQ.ID NO: 6) and Pro-Pro-Gly-Pro-Gln-Gly (SEQ ID NO: 7) and post-translational modifications of the sequences.
110 . The method of claim 109 wherein the three diagnostic sequences have a mass to charge ratio of approximately 301, approximately 471 and approximately 568, respectively.
111 . The method of claim 109 wherein the three diagnostic sequences have a mass to charge ratio of approximately 301, approximately 455 and approximately 552, respectively.Join the waitlist — get patent alerts
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