Shear-enhanced Binding of Intestinal Colonization Factor Antigen l of Enterotoxigenic Escherichia coli
NAVAL MEDICAL RESEARCH CENTER SILVER SPRING MD ENTERIC DISEASES DEPT
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In the intestine, enterotoxigenic Escherichia coli works against peristaltic forces, adhering to the epithelium via the colonization factor antigen I CFA1 fimbria adhesin CfaE. The CfaE adhesin is similar in localization and tertiary but not primary structure to FimH, the type 1 fimbria adhesin of uropathogenic E. coli, which shows shear-dependent binding to epithelial receptors by an allosteric catch-bond mechanism. Thus, we speculated that CfaE is also capable of shear-enhanced binding. Indeed, bovine erythrocytes coursing over immobilized CFA1 fimbriae in flow chambers exhibited low accumulation levels and fast rolling at low shear, but an 80-fold increase in accumulation and threefold decrease in rolling velocity at elevated shear. This effect was reversible and abolished by pre-incubation of fimbriae with anti-CfaE antibody. Erythrocytes bound to whole CfaE in the same shear-enhanced manner, but to CfaE adhesin domain in a shear-inhibitable fashion. Residue replacements designed to disrupt CfaE interdomain interaction decreased the shear dependency of adhesion and increased binding under static conditions to human intestinal epithelial cells. These findings indicate that close interaction between adhesive and anchoring pilin domains of CfaE keeps the former in a low-affinity state that toggles Into a high-affinity state upon separation of two domains, all consistent with an allosteric catch-bond mechanism of CfaE binding.
- Medicine and Medical Research