Evidence for the involvement of essential sulphydryl group in rat hepatic lactogenic receptor but not in somatogenic receptor

Abstract

Incorporation of P-chloromercuribenzene sulphonate (PCMBS) (1mM) in the assay medium for rat hepatic lactogenic receptor produced complete inhibition of binding of [ 125I]oPRL to the membrane. However, the presence of the thiol-reactive agent produced no effect on the binding of [ 125I]bGH to rat hepatic somatogenic receptor. Pretreatment of rat hepatic membrane with PCMBS inhibited the binding of [ 125I]oPRL but not that of [ 125I]bGH. The lactogenic receptor binding inhibition by PCMBS pretreatment was both concentration- and time-dependent, with complete inhibition at 0.5 mM for 60 min at 0°C. Scatchard analysis of [ 125I]oPRL binding to membrane at 50% inhibition by PCMBS (0.11 mM) revealed that the binding capacity was decreased rather than the binding affinity. Furthermore, the inhibition of lactogenic receptor binding by PCMBS could be reversed by dithioerythritol (DTE) treatment. Following 80% inhibition by 0.2 mM PCMBS, full recovery of receptor binding was achieved at 6 mM DTE for 60 min at 0°C. The ‘recovered’ membrane showed no difference from the control membrane in terms of binding capacity and affinity.