| Abstract: | Highly purified microvillus membrane vesicles isolated from rat small intestine were enriched in sucrase, maltase, and aminopeptidase activities. Approximately 90-95% of each enzyme was released from the membrane fraction by treatment with detergent (Triton X-100) and sonication. Using untreated and solubilized preparations, the effect of lectin binding on the activity of each of the three enzymes was measured. It was observed that wheat germ agglutinin (WGA) and phytohemagglutinin (PHA) dramatically enhanced the activity of membrane-bound maltase but had much less effect on the detergent solubilized enzyme. Under the same conditions aminopeptidase activity was inhibited by WGA and PHA while sucrase activity was not affected. These alterations in enzyme activity occurred at lectin concentrations that also precipitated each solubilized enzyme from solution. Inhibitory sugars prevented the alterations in enzyme activity suggesting that the effect is due to the binding of lectin to specific carbohydrate structures. Enhancement of membrane-bound maltase activity by WGA and PHA was shown to be temperature dependent indicating that the lipid environment of the microvillus membrane may play a role in mediating the lectin effect. A kinetic analysis of the changes in maltase activity induced by these two lectins was due solely to an increase in Vmax. Two other lectins used in this study (concanavalin A and Ricinus communis agglutinin) did not readily precipitate the enzymes in question or alter their activity. These results show that binding of lectins to brush border membranes can induce variable changes in the activity of several membrane associated hydrolases, and suggest that similar changes may occur in vivo in the presence of dietary lectin. |
