title: Calcium binding by the PKD1 domain regulates interdomain flexibility in Vibrio cholerae metalloprotease PrtV.
abstract: Vibrio cholerae, the causative agent of cholera, releases several virulence factors including secreted proteases when it infects its host. These factors attack host cell proteins and break down tissue barriers and cellular matrix components such as collagen, laminin, fibronectin, keratin, elastin, and they induce necrotic tissue damage. The secreted protease PrtV constitutes one virulence factors of V. cholerae. It is a metalloprotease belonging to the M6 peptidase family. The protein is expressed as an inactive, multidomain, 102 kDa pre-pro-protein that undergoes several N- and C-terminal modifications after which it is secreted as an intermediate variant of 81 kDa. After secretion from the bacteria, additional proteolytic steps occur to produce the 55 kDa active M6 metalloprotease. The domain arrangement of PrtV is likely to play an important role in these maturation steps, which are known to be regulated by calcium. However, the molecular mechanism by which calcium controls proteolysis is unknown. In this study, we report the atomic resolution crystal structure of the PKD1 domain from V. cholera PrtV (residues 755-838) determined at 1.1 Å. The structure reveals a previously uncharacterized Ca(2+)-binding site located near linker regions between domains. Conformational changes in the Ca(2+)-free and Ca(2+)-bound forms suggest that Ca(2+)-binding at the PKD1 domain controls domain linker flexibility, and plays an important structural role, providing stability to the PrtV protein.
authors.affiliation: Department of Chemistry, Umeå University, Umeå SE-901 87, Sweden ; Umeå Centre for Microbial Research (UCMR), Umeå University, Umeå SE-901 87, Sweden.
authors.forename: Aaron
authors.initials: A
authors.lastname: Edwin
authors.forename: Pramod
authors.initials: P
authors.lastname: Rompikuntal
authors.forename: Erik
authors.initials: E
authors.lastname: Björn
authors.forename: Gunter
authors.initials: G
authors.lastname: Stier
authors.forename: Sun N
authors.initials: SN
authors.lastname: Wai
authors.forename: A Elisabeth
authors.initials: AE
authors.lastname: Sauer-Eriksson
keywords: Atomic resolution; Calcium binding; Metalloprotease; PKD domain; Polycystic Kidney domains; PrtV; Vibrio cholerae; X-ray crystallography
references.citation: Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
references.pmid: 15299374
references.citation: Structure. 2002 Oct;10(10):1453-64
references.pmid: 12377130
references.citation: Cell. 2007 Dec 14;131(6):1047-58
references.pmid: 18083096
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Apr;61(Pt 4):449-57
references.pmid: 15805600
references.citation: Protein Expr Purif. 2009 Mar;64(1):16-23
references.pmid: 18983922
references.citation: Nat Genet. 1995 Jun;10(2):151-60
references.pmid: 7663510
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: Acta Crystallogr D Biol Crystallogr. 2009 Oct;65(Pt 10):1089-97
references.pmid: 19770506
references.citation: Cell. 1995 Apr 21;81(2):289-98
references.pmid: 7736581
references.citation: J Mol Biol. 1990 Oct 5;215(3):403-10
references.pmid: 2231712
references.citation: Microbes Infect. 2000 Jan;2(1):91-8
references.pmid: 10717546
references.citation: Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W197-201
references.pmid: 18463136
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):9280-5
references.pmid: 16754867
references.citation: J Biol Chem. 2012 May 18;287(21):17578-17588
references.pmid: 22442152
references.citation: Biocontrol Sci. 2011 Mar;16(1):1-11
references.pmid: 21467624
references.citation: Bioinformatics. 2008 Dec 1;24(23):2780-1
references.pmid: 18818215
references.citation: Proteins. 2002 May 15;47(3):344-56
references.pmid: 11948788
references.citation: J Appl Microbiol. 1999 Aug;87(2):241-5
references.pmid: 10475957
references.citation: Bioinformatics. 2012 Jun 15;28(12):1658-60
references.pmid: 22556364
references.citation: BMC Microbiol. 2009 Oct 16;9:220
references.pmid: 19835618
references.citation: FEBS J. 2008 Jun;275(12):3167-77
references.pmid: 18479458
references.citation: J Mol Biol. 2007 Sep 21;372(3):774-97
references.pmid: 17681537
references.citation: EMBO J. 1999 Jan 15;18(2):297-305
references.pmid: 9889186
references.citation: Infect Immun. 2001 Oct;69(10):6549-53
references.pmid: 11553605
references.citation: J Biol Chem. 2010 May 7;285(19):14285-91
references.pmid: 20207733
references.citation: Lancet. 2012 Jun 30;379(9835):2466-2476
references.pmid: 22748592
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: J Biol Chem. 2006 Mar 31;281(13):8815-28
references.pmid: 16314409
references.citation: Structure. 2011 Feb 9;19(2):203-11
references.pmid: 21300289
references.citation: J Appl Microbiol. 2005;99(3):551-7
references.pmid: 16108796
references.citation: Microbiol Rev. 1993 Dec;57(4):823-37
references.pmid: 8302217
references.citation: J Biochem. 1997 Feb;121(2):304-8
references.pmid: 9089404
references.citation: J Biol Chem. 2006 Oct 20;281(42):31408-18
references.pmid: 16926147
references.citation: Hum Mol Genet. 2000 Jul 1;9(11):1641-9
references.pmid: 10861291
references.citation: J Mol Biol. 1993 Sep 5;233(1):123-38
references.pmid: 8377180
references.citation: Structure. 2003 Jun;11(6):691-701
references.pmid: 12791257
references.citation: Nat Struct Mol Biol. 2011 Sep 25;18(10):1109-14
references.pmid: 21947205