Expression and purification of the mannose recognition domain of the FimH adhesin.
Type 1 fimbriae have been shown to be specifically required for Escherichia coli colonisation and pathogenesis of the urinary tract. These structural organelles mediate specific adhesion to alpha-D-mannosides by virtue of the FimH adhesin. FimH is a two-domain protein in which the N-terminal domain contains the receptor-binding site and the C-terminal domain is required for organelle integration. To date, FimH has only been isolated as a complex with the system-specific chaperone FimC. Here we report that a functional form of the FimH receptor-binding domain can be readily isolated and characterised by replacing the C-terminal domain with a histidine tag.
Schembri
M A
MA
Department of Microbiology, Bldg 301, Technical University of Denmark, DK-2800, Lyngby, Denmark.
Hasman
H
H
Klemm
P
P
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018830:Adhesins, Escherichia coli; D019677:Artificial Gene Fusion; D001428:Bacterial Vaccines / Q000276:immunology; D015153:Blotting, Western; D002846:Chromatography, Affinity; D017931:DNA Primers; D004926:Escherichia coli / Q000276:immunology / Q000378:metabolism; D022361:Escherichia coli Vaccines; D036781:Fimbriae Proteins; D015870:Gene Expression; D005822:Genetic Vectors; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D018829:Adhesins, Bacterial; D018830:Adhesins, Escherichia coli; D001428:Bacterial Vaccines; D017931:DNA Primers; D022361:Escherichia coli Vaccines; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; C058769:fimH protein, E coli; D036781:Fimbriae Proteins