title: EspP, a serine protease of enterohemorrhagic Escherichia coli, impairs complement activation by cleaving complement factors C3/C3b and C5.
abstract: Hemolytic-uremic syndrome (HUS) is a life-threatening disorder characterized by hemolytic anemia, thrombocytopenia, and renal insufficiency. It is caused mainly by infections with enterohemorrhagic Escherichia coli (EHEC). Recently, Shiga toxin 2, the best-studied virulence factor of EHEC, was reported to interact with complement, implying that complement may be involved in the pathogenesis of EHEC-induced HUS. The aim of the present study was to investigate whether or not the serine protease EspP, an important virulence factor of EHEC, interacts with complement proteins. EspP did not have any effect on the integrity of factor H or factor I. However, EspP was shown to cleave purified C3/C3b and C5. Cleavage of the respective complement proteins also occurred in normal human serum (NHS) as a source of C3/C3b or C5 or when purified complement proteins were added to the supernatant of an EspP-producing wild-type strain. Edman degradation allowed unequivocal mapping of all three main C3b fragments but not of the three main C5 fragments. Complement activation was significantly downregulated in all three pathways for C5-depleted serum to which C5, preincubated with EspP, was added (whereas C5 preincubated with an EspP mutant was able to fully reconstitute complement activation). This indicates that EspP markedly destroyed the functional activity, as measured by a commercial total complement enzyme-linked immunosorbent assay (Wieslab). Downregulation of complement by EspP in vivo may influence the colonization of EHEC bacteria in the gut or the disease severity of HUS.
authors.affiliation: Division of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria. dorothea.orth@i-med.ac.at
authors.forename: Dorothea
authors.initials: D
authors.lastname: Orth
authors.forename: Silvia
authors.initials: S
authors.lastname: Ehrlenbach
authors.forename: Jens
authors.initials: J
authors.lastname: Brockmeyer
authors.forename: Abdul Basit
authors.initials: AB
authors.lastname: Khan
authors.forename: Georg
authors.initials: G
authors.lastname: Huber
authors.forename: Helge
authors.initials: H
authors.lastname: Karch
authors.forename: Bettina
authors.initials: B
authors.lastname: Sarg
authors.forename: Herbert
authors.initials: H
authors.lastname: Lindner
authors.forename: Reinhard
authors.initials: R
authors.lastname: Würzner
mesh_terms: D003167:Complement Activation / Q000187:drug effects*; D003176:Complement C3 / Q000737:chemistry / Q000378:metabolism*; D003179:Complement C3b / Q000737:chemistry / Q000378:metabolism*; D003182:Complement C5 / Q000737:chemistry / Q000378:metabolism*; D054324:Enterohemorrhagic Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D006463:Hemolytic-Uremic Syndrome / Q000382:microbiology; D006801:Humans; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
chemical_list: D003176:Complement C3; D003182:Complement C5; D029968:Escherichia coli Proteins; D003179:Complement C3b; C107189:EspP protein, E coli; D012697:Serine Endopeptidases