CfaE tip mutations in enterotoxigenic Escherichia coli CFA/I fimbriae define critical human intestinal binding sites.
Enterotoxigenic Escherichia coli (ETEC) use colonization factors to attach to the human intestinal mucosa, followed by enterotoxin expression that induces net secretion and diarrhoeal illness. ETEC strain H10407 expresses CFA/I fimbriae, which are composed of multiple CfaB structural subunits and a CfaE tip subunit. Currently, the contribution of these individual fimbrial subunits in intestinal binding remains incompletely defined. To identify the role of CfaE in attachment in the native ETEC background, an R181A single-amino-acid substitution was introduced by recombination into the H10407 genome. The substitution of R181A eliminated haemagglutination and binding of intestinal mucosa biopsies in in vitro organ culture assays, without loss of CFA/I fimbriae expression. Wild-type in trans plasmid-expressed cfaE restored the binding phenotype. In contrast, in trans expression of cfaE containing amino acid 181 substitutions with similar amino acids, lysine, methionine and glutamine did not restore the binding phenotype, indicating that the loss of the binding phenotype was due to localized areas of epitope disruption. R181 appears to have an irreplaceable role in the formation of a receptor-binding feature on CFA/I fimbriae. The results specifically indicate that the CfaE tip protein is a required binding factor in CFA/I-mediated ETEC colonization, making it a potentially important vaccine antigen.
Baker
K K
KK
Center for Vaccine Development, University of Maryland-Baltimore, 685 W. Baltimore St., Baltimore, MD 21201, USA.
Levine
M M
MM
Morison
J
J
Phillips
A
A
Barry
E M
EM
D019943:Amino Acid Substitution; D001422:Bacterial Adhesion / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D036781:Fimbriae Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D010861:Fimbriae, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000382:microbiology; D011995:Recombination, Genetic
C559939:CfaE protein, E coli; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins