Title: Receptor-specific binding of purified F4 to isolated villi
Authors: Van den Broeck, W ×
Cox, E
Goddeeris, Bruno #
Issue Date: 1999
Series Title: Veterinary microbiology vol:68 issue:3-4 pages:255-263
Conference: date:State Univ Ghent, Fac Med Vet, Lab Vet Immunol, B-9820 Merelbeke, Belgium; Katholieke Univ Leuven, Fac Agr & Appl Biol Sci, Lab Physiol & Immunol Domest Anim, B-3001 Heverlee, Belgium
Abstract: Different procedures for preparing and purifying F4ac fimbriae of the enterotoxigenic Escherichia coli strain GIS 26 (O149:K91:F4ac LT(+)Sta(+)STb(+)) were performed and the purity and yield of F4ac were compared. Fimbriae were prepared by either mechanical shearing or heat-shock treatment of concentrated bacterial suspensions (10(11) bacteria/ml). The mechanical shearing procedure resulted in approximately 1.7 mg fimbriae (i.e. 74.4% of the isolated protein) and 0.6 mg (25.6%) contaminating proteins per 10(12) bacteria, whereas the yield of fimbriae following heat-shock treatment was lower (0.3 mg per 10(12) bacteria, i.e. 26.2%) and the relative contamination higher (1.0 mg per 10(12) bacteria, i.e. 73.8%). A further purification consisted of either anion exchange chromatography (AEC) or electro-elution from SDS-polyacrylamide gels. The electroelution procedure was performed under reducing and denaturing conditions, so that purified FaeG subunits, the major subunit of F4, were finally obtained. The binding activity of fimbriae, nonpurified as well as purified, and FaeG to F4-specific receptors on isolated intestinal villi was assessed in an inhibition adhesion assay. Native fimbriae as well as major subunits were able to bind to the receptors, and the specificity of the binding was demonstrated by blockage with F4ac-specific MAb. (C) 1999 Elsevier Science B.V. All rights reserved.
ISSN: 0378-1135
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Division of Gene Technology (-)
× corresponding author
# (joint) last author

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