Subtypes of the plasmid-encoded serine protease EspP in Shiga toxin-producing Escherichia coli: distribution, secretion, and proteolytic activity.
We investigated the prevalence, distribution, and structure of espP in Shiga toxin-producing Escherichia coli (STEC) and assessed the secretion and proteolytic activity of the encoded autotransporter protein EspP (extracellular serine protease, plasmid encoded). espP was identified in 56 of 107 different STEC serotypes. Sequencing of a 3,747-bp region of the 3,900-bp espP gene distinguished four alleles (espPalpha, espPbeta, espPgamma, and espPdelta), with 99.9%, 99.2%, 95.3%, and 95.1% homology, respectively, to espP of E. coli O157:H7 strain EDL933. The espPbeta, espPgamma, and espPdelta genes contained unique insertions and/or clustered point mutations that enabled allele-specific PCRs; these demonstrated the presence of espPalpha, espPbeta, espPgamma, and espPdelta in STEC isolates belonging to 17, 16, 15, and 8 serotypes, respectively. Among four subtypes of EspP encoded by these alleles, EspPalpha (produced by enterohemorrhagic E. coli [EHEC] O157:H7 and the major non-O157 EHEC serotypes) and EspPgamma cleaved pepsin A, human coagulation factor V, and an oligopeptide alanine-alanine-proline-leucine-para-nitroaniline, whereas EspPbeta and EspPdelta either were not secreted or were proteolytically inactive. The lack of proteolysis correlated with point mutations near the active serine protease site. We conclude that espP is widely distributed among STEC strains and displays genetic heterogeneity, which can be used for subtyping and which affects EspP activity. The presence of proteolytically active EspP in EHEC serogroups O157, O26, O111, and O145, which are bona fide human pathogens, suggests that EspP might play a role as an EHEC virulence factor.
Brockmeyer
Jens
J
Institut für Hygiene, Universität Münster, Robert Koch Str. 41, 48149 Münster, Germany. jens.brockmeyer@ukmuenster.de
Bielaszewska
Martina
M
Fruth
Angelika
A
Bonn
Marie Luise
ML
Mellmann
Alexander
A
Humpf
Hans-Ulrich
HU
Karch
Helge
H
D004269:DNA, Bacterial / Q000032:analysis; D003967:Diarrhea / Q000382:microbiology; D054324:Enterohemorrhagic Escherichia coli / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D006463:Hemolytic-Uremic Syndrome / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012697:Serine Endopeptidases / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D012703:Serotyping; D022621:Shiga Toxin / Q000096:biosynthesis*
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D022621:Shiga Toxin; C107189:EspP protein, E coli; D012697:Serine Endopeptidases
Mol Membr Biol. 1997 Jul-Sep;14(3):113-23
9394291
Proc Natl Acad Sci U S A. 2005 Jan 4;102(1):221-6
15615856
Clin Infect Dis. 2005 Sep 15;41(6):785-92
16107974
J Infect Dis. 2005 Oct 15;192(8):1422-9
16170761
Int J Med Microbiol. 2005 Oct;295(6-7):405-18
16238016
Mol Microbiol. 2005 Nov;58(4):945-58
16262782
Mol Biol Evol. 2006 Feb;23(2):254-67
16221896
J Biol Chem. 2006 Apr 7;281(14):9038-48
16455668
Clin Infect Dis. 2006 Nov 1;43(9):1168-9
17029136
Clin Infect Dis. 2006 Dec 15;43(12):1587-95
17109294
Appl Environ Microbiol. 2007 Mar;73(5):1553-62
17220264
EMBO J. 2007 Apr 4;26(7):1942-52
17347646
FEMS Microbiol Lett. 2007 Jun;271(2):258-64
17451446
Lancet. 2005 Mar 19-25;365(9464):1073-86
15781103
Infect Immun. 2001 Jan;69(1):315-24
11119520
Infect Immun. 2001 Mar;69(3):1231-43
11179284
J Bacteriol. 2001 Aug;183(15):4626-35
11443098
Arch Dis Child. 2001 Aug;85(2):125-31
11466187
Pediatr Res. 2001 Aug;50(2):163-71
11477199
Infect Immun. 2001 Nov;69(11):6863-73
11598060
J Infect Dis. 2002 Aug 15;186(4):493-500
12195376
Infect Immun. 2002 Dec;70(12):7105-13
12438392
J Bacteriol. 2003 Mar;185(6):1831-40
12618447
Int J Med Microbiol. 2003 Feb;292(7-8):477-86
12635930
J Infect Dis. 2003 Sep 1;188(5):719-29
12934188
JAMA. 2003 Sep 10;290(10):1379-81
12966132
Infect Immun. 2003 Nov;71(11):6307-19
14573650
Mol Biotechnol. 2004 Feb;26(2):117-22
14764937
Res Microbiol. 2004 Mar;155(2):53-60
14990256
J Clin Microbiol. 2004 Mar;42(3):1099-108
15004060
Nat Rev Microbiol. 2004 Feb;2(2):123-40
15040260
J Biol Chem. 2004 Jul 23;279(30):31495-504
15151995
Brief Bioinform. 2004 Jun;5(2):150-63
15260895
J Bacteriol. 2004 Aug;186(16):5432-41
15292145
Nature. 1970 Aug 15;227(5259):680-5
5432063
Nature. 1987 Jan 29-Feb 4;325(6103):458-62
3027577
J Infect Dis. 1987 Jul;156(1):175-82
3298451
Gastroenterology. 1990 Jul;99(1):142-9
2188868
Infect Immun. 1992 Jan;60(1):13-8
1729177
Mol Phylogenet Evol. 1992 Sep;1(3):242-52
1342941
Infect Immun. 1994 Aug;62(8):3463-71
8039917
J Infect Dis. 1997 Jun;175(6):1382-9
9180177
Mol Microbiol. 1997 May;24(4):767-78
9194704
Mol Microbiol. 1997 Aug;25(4):771-84
9379905
Trends Microbiol. 1998 Sep;6(9):370-8
9778731
J Clin Microbiol. 1999 Mar;37(3):497-503
9986802
Infect Immun. 1999 May;67(5):2184-92
10225873
Microbiology. 1999 May;145 ( Pt 5):1005-14
10376815
J Clin Microbiol. 1999 Oct;37(10):3357-61
10488206
J Clin Microbiol. 1999 Nov;37(11):3491-6
10523540
Mol Nutr Food Res. 2004 Dec;48(7):504-14
15538707
Microbiol Mol Biol Rev. 2004 Dec;68(4):692-744
15590781
J Clin Microbiol. 2005 Aug;43(8):4225-8
16081985