Two specific sites for binding of K88ab Escherichia coli fimbriae to porcine intestinal brush border membranes.

Abstract

We have studied the characteristics of the binding of the K88ab Escherichia coli fimbrial antigen to porcine brush border membranes by solid phase binding assay. Binding of biotinylated K88ab to brush border membranes followed a sigmoidal dependence and was saturable, apparent saturation occurring with 0.8 ng of fimbriae (approx. 7 ng of fimbriae per microg of brush border protein) irrespective of incubation temperature in the range of room temperature to 4 degrees C. A Hill plot of log [(fimbriae bound)/(maximal binding-fimbriae bound)] vs. log free fimbriae gave a maximal slope of about 2, indicating the existence of two binding sites. From an analysis of an Scatchard plot, apparent binding constants (1)K(2) and (2)K(2) of 7.1 x 10(8) and 17.1 x 10(8)M(-1) were obtained at room temperature. Nor did temperature have any effect on the rate of binding or on receptor affinity (S(0.5)).