Regulation of Na+-H+ exchange by transmethylation reactions in rat colonic brush-border membranes.

Incubation of rat colonic brush-border membrane vesicles with 200 microM S-adenosyl-L-[Me-3H]methionine resulted in the labeling of both membrane phospholipids and proteins. This labeling was decreased approximately 50% by the methylation inhibitor S-adenosyl-L-homocysteine (2 mM). Utilizing the pH-sensitive fluorescent dye, acridine orange, as a means of determining Na+-H+ exchange, S-adenosyl-L-methionine (200 microM) significantly increased sodium-stimulated proton efflux in these vesicles at all concentrations of sodium (2.5-50 mM) tested. Examination of the kinetic parameters for sodium-stimulated proton efflux in the presence and absence of 200 microM S-adenosyl-L-methionine revealed that the methyl donor increased the Vmax for this exchange mechanism (expressed in arbitrary fluorescence units) by approx. 36% but did not influence its Km for sodium. S-Adenosyl-L-homocysteine (2 mM) inhibited S-adenosyl-L-methionine-mediated stimulation of this exchange process. The results demonstrate that methylation of membrane phospholipids and/or proteins can modulate Na+-H+ exchange in rat colonic brush-border membrane vesicles.