Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes.
We examined the effect of brefeldin A, an antiviral antibiotic, on protein synthesis, intracellular processing, and secretion in primary culture of rat hepatocytes. The secretion was strongly blocked by the drug at 1 microgram/ml and higher concentrations, while the protein synthesis was maintained fairly well. Pulse-chase experiments with [35S]methionine demonstrated that brefeldin A completely blocked the proteolytic conversion of proalbumin to serum albumin up to 60 min of chase, although its conversion was observed as early as 20 min in the control cells. The drug also inhibited the terminal glycosylation of oligosaccharide chains of alpha 1-protease inhibitor and haptoglobin. These two modifications have been shown to occur at the trans region of the Golgi complex. The drug, however, had no effect on the proteolytic processing of the haptoglobin proform which takes place within the endoplasmic reticulum. Such an effect by brefeldin A is very similar with that induced by the carboxylic ionophore monensin. However, in contrast to evidence that monensin causes a delayed secretion of the unprocessed forms of these proteins, brefeldin A allowed the completely processed forms to be secreted after a prolonged accumulation of the unprocessed forms. Morphological observations demonstrated that the endoplasmic reticulum was markedly dilated by treatment with the drug at 10 micrograms/ml which continuously blocked the secretion. On the other hand, brefeldin A caused no inhibitory effect on the endocytic pathway as judged by cellular uptake and degradation of 125I-asialofetuin. These results indicate that brefeldin A is a unique agent which primarily impedes protein transport from the endoplasmic reticulum to the Golgi complex by a mechanism different from those considered for other secretion-blocking agents so far reported.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001212:Asialoglycoproteins; D001798:Blood Proteins / Q000378:metabolism; D001826:Body Fluids / Q000378:metabolism*; D020126:Brefeldin A; D050260:Carbohydrate Metabolism; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology*; D004705:Endocytosis; D060748:Fetuins; D006242:Haptoglobins / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008854:Microscopy, Electron; D011228:Prealbumin / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D013997:Time Factors; D000515:alpha 1-Antitrypsin; D000509:alpha-Fetoproteins / Q000378:metabolism
D001212:Asialoglycoproteins; D001798:Blood Proteins; D003517:Cyclopentanes; D060748:Fetuins; D006242:Haptoglobins; D011228:Prealbumin; D011506:Proteins; D000515:alpha 1-Antitrypsin; D000509:alpha-Fetoproteins; C017029:asialofetuin; D020126:Brefeldin A; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000255:Adenosine Triphosphate