Type 1 fimbrial shafts of Escherichia coli and Klebsiella pneumoniae influence sugar-binding specificities of their FimH adhesins.
The type 1 fimbriae of enterobacteria comprise FimA, which constitutes most of the fimbrial shaft, and a cassette of three minor ancillary subunits including FimH, the mannose-binding moiety. The sugar-binding specificities of Escherichia coli and Klebsiella pneumoniae type 1 fimbriae were examined by determining the relative activities of two aromatic mannosides in inhibiting the yeast aggregation caused by the fimbriated bacteria. 4-Methylumbelliferyl alpha-mannoside (MeUmb alpha Man) was approximately 10-fold more effective than p-nitrophenyl alpha-mannoside (p-NP alpha Man) in inhibiting the yeast aggregation caused by the recombinant expressing native E. coli type 1 fimbriae. In contrast, MeUmb alpha Man was only fourfold more effective than p-NP alpha Man in assays employing the recombinant expressing native K. pneumoniae type 1 fimbriae. In order to elucidate the molecular mechanisms underlying the sugar-binding specificities of type 1 fimbriae in the two species, transcomplementation studies were performed and resulted in the creation of recombinants expressing two types of hybrid fimbriae: one consisting of a cassette of minor subunits of E. coli fimbriae borne on a filamentous shaft of K. pneumoniae FimA subunits and the other consisting of a cassette of K. pneumoniae minor fimbrial subunits borne on a shaft of E. coli FimA subunits. Although the heterologous FimH was incorporated into the fimbrial filaments in amounts comparable to those observed in native fimbriae, the hemagglutination activities of recombinants expressing hybrid fimbriae were significantly lower than those of their counterparts bearing native fimbriae. The sugar-binding specificity of the recombinant expressing hybrid fimbriae consisting of an E. coli shaft bearing K. pneumoniae FimH was different from those of recombinants expressing native K. pneumoniae fimbriae in its affinity for the two aromatic sugars but was remarkably similar to the specificities exhibited by recombinants expressing native E. coli fimbriae. Conversely, the sugar-binding specificity of the recombinant expressing hybrid fimbriae consisting of a K. pneumoniae shaft bearing E. coli FimH was different from that of the recombinant expressing native E. coli fimbriae but was very similar to those of recombinants expressing native K. pneumoniae fimbriae. We conclude that the differences in the sugar-binding specificity between E. coli and K. pneumoniae FimH fimbrial subunits is influenced by the fimbrial shafts which carry the adhesin molecules in a functionally competent form at the distal tips.
Madison
B
B
Department of Clinical Laboratory Sciences, University of Tennessee, Memphis 68318.
Ofek
I
I
Clegg
S
S
Abraham
S N
SN
D018830:Adhesins, Escherichia coli; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D004926:Escherichia coli / Q000502:physiology*; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000502:physiology*; D007711:Klebsiella pneumoniae / Q000502:physiology*
D018830:Adhesins, Escherichia coli; D001426:Bacterial Proteins; C058769:fimH protein, E coli; D036781:Fimbriae Proteins