Crystal structure of Spy0129, a Streptococcus pyogenes class B sortase involved in pilus assembly.
Sortase enzymes are cysteine transpeptidases that mediate the covalent attachment of substrate proteins to the cell walls of gram-positive bacteria, and thereby play a crucial role in virulence, infection and colonisation by pathogens. Many cell-surface proteins are anchored by the housekeeping sortase SrtA but other more specialised sortases exist that attach sub-sets of proteins or function in pilus assembly. The sortase Spy0129, or SrtC1, from the M1 SF370 strain of Streptococcus pyogenes is responsible for generating the covalent linkages between the pilin subunits in the pili of this organism. The crystal structure of Spy0129 has been determined at 2.3 Å resolution (R = 20.4%, Rfree  = 26.0%). The structure shows that Spy0129 is a class B sortase, in contrast to other characterised pilin polymerases, which belong to class C. Spy0129 lacks a flap believed to function in substrate recognition in class C enzymes and instead has an elaborated β6/β7 loop. The two independent Spy0129 molecules in the crystal show differences in the positions and orientations of the catalytic Cys and His residues, Cys221 and His126, correlated with movements of the β7/β8 and β4/β5 loops that respectively follow these residues. Bound zinc ions stabilise these alternative conformations in the crystal. This conformational variability is likely to be important for function although there is no evidence that zinc is involved in vivo.
Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, New Zealand.
Hae Joo
HJ
Kang
Fasséli
F
Coulibaly
Thomas
T
Proft
Edward N
EN
Baker
D000595:Amino Acid Sequence; D000596:Amino Acids; D019881:Aminoacyltransferases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D036781:Fimbriae Proteins / Q000737:chemistry; D010861:Fimbriae, Bacterial / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding; D016415:Sequence Alignment; D013297:Streptococcus pyogenes / Q000201:enzymology*; D040681:Structural Homology, Protein; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry
D000596:Amino Acids; D001426:Bacterial Proteins; C509821:sortase B; D036781:Fimbriae Proteins; D019881:Aminoacyltransferases; D003546:Cysteine Endopeptidases; D015032:Zinc
N Engl J Med. 1996 Aug 22;335(8):590-1
8678939
Infect Immun. 2002 Mar;70(3):1159-67
11854196
Res Microbiol. 2005 Apr;156(3):289-97
15808931
Science. 1999 Jul 30;285(5428):760-3
10427003
Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2293-8
11830639
J Biol Chem. 2000 Mar 31;275(13):9876-81
10734144
J Bacteriol. 2010 Sep;192(18):4651-9
20639332
Acta Crystallogr D Biol Crystallogr. 1996 Jan 1;52(Pt 1):30-42
15299723
Methods Mol Biol. 2007;364:215-30
17172768
J Biol Chem. 2009 Mar 13;284(11):6924-33
19129180
Structure. 2004 Jul;12(7):1147-56
15242591
J Biol Chem. 2002 May 3;277(18):16241-8
11856734
Nucleic Acids Res. 1988 Aug 11;16(15):7351-67
3045756
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68
15572779
Infect Immun. 2004 May;72(5):2710-22
15102780
J Biol Chem. 2009 Sep 4;284(36):24465-77
19592495
Proc Natl Acad Sci U S A. 2001 May 22;98(11):6056-61
11371637
Mol Microbiol. 2007 Nov;66(4):961-74
17919283
Mol Microbiol. 2003 Nov;50(4):1429-38
14622427
Pharmacol Rev. 2008 Mar;60(1):128-41
18321961
Mol Microbiol. 2006 Oct;62(2):320-30
16978260
Mol Microbiol. 2006 Jun;60(6):1401-13
16796677
Nat Rev Microbiol. 2006 Jul;4(7):509-19
16778837
Methods Enzymol. 1997;276:307-26
27754618
Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15641-6
16223875
Structure. 2008 Dec 10;16(12):1838-48
19081060
Science. 2007 Dec 7;318(5856):1625-8
18063798
Acta Crystallogr D Biol Crystallogr. 2001 Jan;57(Pt 1):122-33
11134934
J Biol Chem. 2004 Jul 23;279(30):31383-9
15117963
Infect Immun. 2003 May;71(5):2758-65
12704150
J Biol Chem. 2007 Mar 2;282(9):6571-81
17200112
J Biol Chem. 2010 Jul 23;285(30):23433-43
20489200
FEBS Lett. 2004 Jul 30;571(1-3):221-6
15280046
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2857-62
16481624
J Biol Chem. 2002 Mar 1;277(9):7447-52
11714722
J Bacteriol. 2007 Feb;189(4):1426-34
17012387
J Biol Chem. 2010 Jun 25;285(26):20381-9
20427291
J Biol Chem. 2006 Jan 20;281(3):1817-26
16269411
J Biol Chem. 2010 Oct 29;285(44):33858-66
20729215
J Biol Chem. 2004 Sep 3;279(36):37763-70
15247224
Mol Microbiol. 2001 Jun;40(5):1049-57
11401711
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
J Bacteriol. 2008 Jan;190(2):527-35
17993527
Infect Immun. 2003 Mar;71(3):1361-9
12595453
Structure. 2004 Jan;12(1):105-12
14725770
Biochemistry. 2007 Jun 19;46(24):7269-78
17518446
Mini Rev Med Chem. 2007 Oct;7(10):991-1000
17979801
Cell. 1992 Jul 24;70(2):267-81
1638631
Cell Microbiol. 2007 Jul;9(7):1822-33
17359232
Mol Microbiol. 2007 May;64(4):968-83
17501921
EMBO J. 1993 Dec;12(12):4803-11
8223489
Mol Microbiol. 1994 Oct;14(1):115-21
7830549
Proc Natl Acad Sci U S A. 2000 May 9;97(10):5510-5
10805806
Anal Biochem. 2004 Mar 1;326(1):42-8
14769334
J Infect Dis. 2002 May 15;185(10):1417-24
11992276
J Bacteriol. 2006 Dec;188(23):8145-52
17012401
Proc Natl Acad Sci U S A. 2009 Oct 6;106(40):16967-71
19805181
Protein Expr Purif. 2005 May;41(1):207-34
15915565
Nat Struct Biol. 1999 May;6(5):458-63
10331874
J Mol Biol. 2009 Oct 30;393(3):704-16
19729023