Biogenesis of microsomal membrane glycoproteins in rat liver. III. Release of glycoproteins from the Golgi fraction and their transfer to microsomal membranes.
The presence in the Golgi fraction of glycoproteins destined to be incorporated into the microsomal membrane was investigated. When incubated in sucrose, washed Golgi vesicles released four major, weakly acidic glycoproteins, some of which could be incorporated into microsomal membranes by incubation. Double labeling with [3H]glucosamine and [14C]leucine demonstrated the incorporation of both protein and oligosaccharide moieties, and the main peak of radioactivity was associated with the 70,000 mol wt region after SDS-gel electrophoresis. The proteins that could be incorporated into microsomes were probably associated to a large extent with the outer surface of the Golgi membrane. Centrifugation of the proteins released from the Golgi in a KBr solution (p = 1.24) resulted in a separation of glycoproteins, those in the top layer most actively incorporated into microsomes. The lipoglycoproteins in the top layer that could be incorporated appeared in the 70,000 mol wt region after SDS-gel electrophoresis, as did the corresponding proteins isolated from the supernate. These results suggest that glycoproteins with completed oligosaccharide chains are released from the Golgi system to the cytosol and are subsequently transferred to microsomes as constitutive membrane components.
D000818:Animals; D000906:Antibodies; D002458:Cell Fractionation; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D006056:Golgi Apparatus / Q000032:analysis / Q000378:metabolism*; D007118:Immunoassay; D008099:Liver / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008566:Membranes / Q000378:metabolism; D008862:Microsomes, Liver / Q000032:analysis / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D008970:Molecular Weight; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
D000906:Antibodies; D006023:Glycoproteins