Linker insertion analysis of the FimH adhesin of type 1 fimbriae in an Escherichia coli fimH-null background.
The gene encoding the Escherichia coli FimH adhesin of type 1 fimbriae has been subjected to linker insertion mutagenesis. Amino acid changes were introduced at a number of positions spanning the entire sequence in order to probe the structure-function relationship of the FimH protein. The effect of these mutations on the ability of bacteria to express a D-mannose binding phenotype was assessed in a fimH null mutant (MS4) constructed by allelic exchange in the E. coli K-12 strain PC31. Mutations mapping at amino acid residues 36, 58 and 279 of the mature FimH protein were shown to completely abolish binding to D-mannose receptors. Differences in the level of fimbriation were also observed as a result of some of the mutations in the fimH gene. These mutants may prove useful in dissecting receptor-ligand interactions by defining regions of the FimH protein that are important in erythrocyte binding.
Schembri
M A
MA
Department of Microbiology, Technical University of Denmark, Lyngby, Denmark.
Pallesen
L
L
Connell
H
H
Hasty
D L
DL
Klemm
P
P
D018829:Adhesins, Bacterial / Q000235:genetics* / Q000502:physiology; D018830:Adhesins, Escherichia coli; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure; D005798:Genes, Bacterial; D006168:Guinea Pigs; D006384:Hemagglutination; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D010641:Phenotype
D018829:Adhesins, Bacterial; D018830:Adhesins, Escherichia coli; D017931:DNA Primers; C058769:fimH protein, E coli; D036781:Fimbriae Proteins