Sulfhydryl groups essential for the volume-sensitive release of taurine from astrocytes.

Abstract

The volume-sensitive [3H]taurine efflux from cultured astrocytes was found to be strongly inhibited by the sulfhydryl group-modifying reagent N-ethylmaleimide (NEM). This maleimide inhibited taurine efflux evoked by 50% hyposmotic solutions with an IC50 of 54 microM. The inhibition by NEM followed pseudo-first order reaction kinetics. A double log plot of the pseudo-first order constant against NEM concentration gave a linear relationship with a slope of 1.2. The data are consistent with a simple bimolecular reaction mechanism in which one molecule of NEM reacts with one sulfhydryl group per transport unit. The membrane-impermeant maleimide derivative 5-eosin maleimide did not affect the volume-stimulated taurine efflux. The sulfhydryl-modifying mercurial reagents mersalyl and p-chloro mercuribenzenesulfonate (0.5-1 mM) increased [3H]taurine efflux under isosmotic conditions and concomitantly decreased the hyposmolarity-evoked efflux. The results demonstrate an essential requirement for sulfhydryl groups for the volume-sensitive taurine efflux.