Transport of the vesicular stomatitis glycoprotein to trans Golgi membranes in a cell-free system.
Terminal steps in the transport of the vesicular stomatitis virus glycoprotein (G protein) in the Golgi stack have been reconstituted in a cell-free system. Incorporation of sialic acid into the oligosaccharide chains of G protein was used to monitor transport into the trans Golgi compartment. Transport-coupled sialylation required cytosol, ATP, an N-ethylmaleimide-sensitive factor extractable from Golgi membranes, and long chain acyl coenzyme A. The G protein receiving sialic acid in the cell-free system begins its in vitro transport bearing galactose residues acquired in vivo. Earlier reports (Balch, W. E., Dunphy, W. G., Braell, W. A., and Rothman, J. E. (1984a) Cell 39, 405-416) documented that transport of G protein into the medial (GlcNAc Transferase-containing) compartment is reconstituted under the same conditions. On the basis of the results reported here, it now appears that a more complete set of transport operations of the Golgi stack may be simultaneously reconstituted.
D000214:Acyl Coenzyme A / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002474:Cell-Free System; D006224:Cricetinae; D003412:Cricetulus; D003600:Cytosol / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis; D006056:Golgi Apparatus / Q000378:metabolism*; D008562:Membrane Glycoproteins; D019158:N-Acetylneuraminic Acid; D010053:Ovary; D011499:Protein Processing, Post-Translational; D012794:Sialic Acids / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism*; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000378:metabolism*
D000214:Acyl Coenzyme A; C028391:G protein, vesicular stomatitis virus; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D012794:Sialic Acids; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D019158:N-Acetylneuraminic Acid; D005033:Ethylmaleimide; D005690:Galactose