title: Subtypes of the plasmid-encoded serine protease EspP in Shiga toxin-producing Escherichia coli: distribution, secretion, and proteolytic activity.
abstract: 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.
authors: Institut für Hygiene, Universität Münster, Robert Koch Str. 41, 48149 Münster, Germany. jens.brockmeyer@ukmuenster.de
authors: Jens
authors: J
authors: Brockmeyer
authors: Martina
authors: M
authors: Bielaszewska
authors: Angelika
authors: A
authors: Fruth
authors: Marie Luise
authors: ML
authors: Bonn
authors: Alexander
authors: A
authors: Mellmann
authors: Hans-Ulrich
authors: HU
authors: Humpf
authors: Helge
authors: H
authors: Karch
mesh_terms: 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*
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D022621:Shiga Toxin; C107189:EspP protein, E coli; D012697:Serine Endopeptidases
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