Inhibition by brefeldin A of NADH oxidation activity of rat liver Golgi apparatus accelerated by GDP

Abstract

Al~traetReduced pyridine nucleotide has been reported to enhance cell-free transfer of membrane material from a radiolabeled Golgi apparatus donor fraction from rat liver to an acceptor fraction consisting of inside-out plasma vesicles immobilized on nitrocellulose [(1992) Bioehim. Biophys. Acta 1107, 131]. As part of a continuing effort to identify NADH-requiring enzymes in the Goigi apparatus which may be important to membrane trafficking, highly purified fractions of Golgi apparatus from rat liver were tested for their ability to oxidize NADH and the inhibition of the oxidation of NADH by brefeldin A. The isolated Golgi apparatus fractions were found to oxidize NADH with a specific activity comparable to that of the plasma membrane of rat liver. The activity was inhibited by brefeldin A and this inhibition was augmented by GDP. At near optimal concentrations of 7/tM brefeldin A and 1/aM GDP, the activity was > 90% inhibited. Brefeldin A inhibition of NADH oxidation by the Golgi apparatus was time-dependent and GDP appeared to accelerate the inhibition by brefeidin A.