US2013144038A1PendingUtilityA1

CRYSTAL STRUCTURE OF A MarR FAMILY POLYPEPTIDE

Individually held — no corporate assignee on recordPriority: Jul 13, 2001Filed: May 15, 2012Published: Jun 6, 2013
Est. expiryJul 13, 2021(expired)· nominal 20-yr term from priority
C07K 2299/00C07K 14/245G01N 23/20
50
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Claims

Abstract

The crystal structure of the product, crystals of the MarR protein, a regulator of multiple antibiotic resistance in Escherichia coli, and methods of crystallization of the MarR protein are described.

Claims

exact text as granted — not AI-modified
1 . A crystallized MarR family polypeptide, wherein said crystallized MarR family polypeptide is crystallized under appropriate conditions such that the three dimensional structure of said MarR family polypeptide can be determined. 
     
     
         2 . The crystallized MarR family polypeptide of  claim 1 , wherein the crystallized MarR family polypeptide is a structural homolog of crystallized MarR. 
     
     
         3 . The crystallized MarR family polypeptide of  claim 1 , wherein said MarR family polypeptide is selected from the group consisting of MarR, SlyA, EmrR (MprA), PapX, PrsX, HpcR, Ec17kD of  Escherichia coli;  MarR, SlyA, EmrR of  Salmonella typhimurium;  MexR of  Pseudomonas aeruginosa;  PecS of  Erwinia chrysanthemi;  BadR of  Rhodopseudomonas palustris;  OrfE of  Burkholderia pseudomallei;  YdcH, YhbI, YkmA, YkoM, Orf7, YfiV, YetL, YdgJ, YwoH, YwaE, YwhA, Hpr, YybA, YxaD, YsmB, YusO, YpoP, YkvE of  Bacillus subtilus;:  Orf7 of  Bacillus firmus;:  Orf145, Orf141 of  Staphylococcus sciuri;:  CinR of  Butyrivibrio fibrisolvens;:  Orf158 of  Sphingomonas aromaticivorans;:  NhhD of  Rhodococcus rhodochrous;  Orf1 of  Streptomyces peucetius;:  14.7 kD, Rv1404, Rv0737, Rv0042c, Yz08 (15.6 kD) of  Mycobacterium tuberculosis;  Yz08 (15.6 kD) of  Mycobacterium leprae;:  MTH313 of  Methanobacterium thermoautotrophicum;  Lrs14 of  Sulfolobus solfataricus;  CinR of  Archaeoglobus fulgidus;  PetP of  Rhodobacter capsulatus;  and SlyA (E293909) of  Sinorhizobium meliloti.    
     
     
         4 . The crystallized MarR family polypeptide of  claim 1 , wherein said MarR family polypeptide is MarR. 
     
     
         5 . The crystallized MarR family polypeptide of  claim 4 , wherein the peptide sequence of MarR comprises the amino acid sequence shown in SEQ ID NO:2. 
     
     
         6 . The crystallized MarR family polypeptide of  claim 4 , wherein said three dimensional structure has the space group of I4 1 22. 
     
     
         7 . The crystallized MarR family polypeptide of  claim 1 , wherein said appropriate conditions include the presence of a MarR family polypeptide modulator. 
     
     
         8 . The crystallized MarR family polypeptide of  claim 7 , wherein said MarR family polypeptide inhibitor is salicylate. 
     
     
         9 . The crystallized MarR family polypeptide of  claim 1 , wherein said three dimensional structure is determined to a resolution of 5 Å or better. 
     
     
         10 .- 12 . (canceled) 
     
     
         13 . The crystallized MarR family polypeptide of  claim 4 , wherein said crystallized MarR family polypeptide has a structure defined by the atomic coordinates given in  FIG. 7  or  FIG. 8 . 
     
     
         14 . (canceled) 
     
     
         15 . The crystallized MarR family polypeptide of  claim 1 , wherein said MarR family polypeptide has a winged-helix structure. 
     
     
         16 . A crystallized MarR family polypeptide defined by the atomic coordinates given in  FIG. 7  or  FIG. 8 . 
     
     
         17 . (canceled) 
     
     
         18 . A method for determining the three dimensional structure of MarR family polypeptide, comprising
 crystallizing the MarR family polypeptide under appropriate conditions such that crystals are formed; and   analyzing the crystallized MarR family polypeptide, such that the three dimensional structure of the MarR family polypeptide is determined.   
     
     
         19 . The method of  claim 18 , wherein said crystallized MarR family polypeptide is analyzed by x-ray diffraction techniques. 
     
     
         20 . The method of  claim 18 , wherein said MarR family polypeptide is MarR. 
     
     
         21 . The method of  claim 20 , wherein MarR comprises the amino acid sequence set forth in SEQ ID NO:2. 
     
     
         22 . The method of  claim 18 , wherein said appropriate conditions comprise the presence of a MarR family polypeptide modulator. 
     
     
         23 . The method of  claim 22 , wherein said MarR family polypeptide modulator is salicylate. 
     
     
         24 . The method of  claim 18 , wherein said three dimensional structure is determined to a resolution of 5 Å or better. 
     
     
         25 .- 27 . (canceled) 
     
     
         28 . The method of  claim 18 , wherein said MarR family polypeptide is selected from the group consisting of: MarR, SlyA, EmrR (MprA), PapX, PrsX, HpcR, Ec17kD of  Escherichia coli;  MarR, SlyA, EmrR of  Salmonella typhimurium;  MexR of  Pseudomonas aeruginosa;  PecS of  Erwinia chrysanthemi;  BadR of  Rhodopseudomonas palustris;  OrfE of  Burkholderia pseudomallei;  YdcH, YhbI, YkmA, YkoM, Orf7, YfiV, YetL, YdgJ, YwoH, YwaE, YwhA, Hpr, YybA, YxaD, YsmB, YusO, YpoP, YkvE of  Bacillus subtilus;:  Orf7 of  Bacillus firmus;:  Orf145, Orf141 of  Staphylococcus sciuri;:  CinR of  Butyrivibrio fibrisolvens;:  Orf158 of  Sphingomonas aromaticivorans;:  NhhD of  Rhodococcus rhodochrous;  Orf1 of  Streptomyces peucetius;:  14.7 kD, Ry1404, Rv0737, Rv0042c, Yz08 (15.6 kD) of  Mycobacterium tuberculosis;  Yz08 (15.6 kD) of  Mycobacterium leprae;:  MTH313 of  Methanobacterium thermoautotrophicum;  Lrs14 of  Sulfolobus solfataricus;  CinR of  Archaeoglobus fulgidus;  PetP of  Rhodobacter capsulatus;  and SlyA (E293909) of  Sinorhizobium meliloti.    
     
     
         29 . (canceled)

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