PMID: 6410180Jan 1, 1983Paper

Soluble pig intestinal cell membrane components with affinities for E. coli K88+ antigen

Molecular and Cellular Biochemistry
T E Staley, I B Wilson

Abstract

Pig intestinal brush borders (BB) were radiolabeled by iodination using the lactoperoxidase-hydrogen peroxide procedure. The BB were then detergent solubilized, centrifuged to remove particulate material, and chromatographed on Sepharose CL-4B. The fractions were incubated with K88+ E. coli using an in vitro binding assay. Binding of the iodinated membranes to K88+ E. coli occurred throughout a wide range of molecular weight components, in excess of 690K daltons to near 25K daltons. The system utilizing intact K88+ E. coli and solubilized BB was shown to be saturable. Prior contact of K88+ E. coli with nonradiolabeled membranes or specific antibodies to K88+ pili inhibited binding of the radiolabeled BB. Simple sugars were tested for their ability to block binding of the labeled BB; partial inhibition occurred with galactose (17.9%), galactosamine (32%), glucose (10.6%), and N-acetylglucosamine (32%). Calcium enhanced binding with as little as 10 microM. A 10 x increase in binding occurred with 500 microM calcium. Affinity chromatography using K88+ pili coupled on agarose beads avidly bound the labeled BB. The receptor membranes were eluted with high molar concentrations of salt, however considerable degradation occurred. Despi...Continue Reading

Citations

Oct 6, 1999·Veterinary Microbiology·M D JeyasinghamD Kelly
May 1, 1992·Microbial Pathogenesis·P T Willemsen, F K de Graaf
Mar 6, 2007·Comparative Immunology, Microbiology and Infectious Diseases·Maria J Caloca, Susana Suarez

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