J. Biol. Chem., Vol. 265, Issue 3, 1470-1475, 01, 1990

Hormone-induced system A amino acid transport activity in rat liver plasma membrane and Golgi vesicles. Evidence for a differential sensitivity to inactivation by N-ethylmaleimide during carrier maturation

R Cariappa and MS Kilberg
Department of Biochemistry and Molecular Biology, J. Hillis Miller Health Center, University of Florida, Gainesville 32610.

We have investigated the biogenesis and processing of the rat hepatic System A amino acid carrier following induction of its de novo synthesis by the combined action of glucagon and dexamethasone. Golgi subfractions isolated from hormone-treated rat liver form transport competent vesicles and possess characteristic System A activity based on pH sensitivity and 2-(methylamino)isobutyric acid inhibition of Na(+)-dependent 2-aminoisobutyric acid uptake. We have monitored the time course for appearance of the newly synthesized carrier in the Golgi and plasma membrane fractions after the administration of hormones. Our data suggest that it may also be possible to detect processing intermediates of the System A carrier in the Golgi. Perfusion of whole rat liver with 5 mM N-ethylmaleimide followed by isolation of Golgi subfractions and plasma membrane revealed a differential sensitivity such that the plasma membrane or trans Golgi activities were inactivated to a much greater extent than those of the cis or medial Golgi. In vitro N-ethylmaleimide treatment of membrane fractions isolated from an intact rat results in an inactivation of the trans Golgi and plasma membrane System A carrier protein, whereas the cis and medial Golgi fractions retained their transport activity.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?