The glycoprotein that is transported between successive compartments of the Golgi in a cell-free system resides in stacks of cisternae.
Electron microscope autoradiography has been used to localize the glycoprotein transported between successive compartments of the Golgi in a cell-free system. Both donor and acceptor Golgi fractions contain stacks of cisternae, which remain as discrete populations even after prolonged incubations together. The glycosylated VSV G protein, having received 3H-GlcNAc residues following its transport in vitro, is entirely within the population of acceptor stacks from the uninfected wild-type Golgi population (those housing GlcNAc transferase l). Quantitation of 3H grains reveals between 6,000 and 12,000 molecules of G protein introduced into each acceptor cisterna as a result of transport in the cell-free system, amounting to approximately 5% of its total membrane protein. This represents about the full complement of transported protein normally contained in a Golgi cisterna in vivo. Transport in the cell-free system is efficient and specific, preserving the integrity of the Golgi stack and its individual cisternae.
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001345:Autoradiography; D001692:Biological Transport; D002460:Cell Line; D002472:Cell Transformation, Viral; D002474:Cell-Free System; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D008562:Membrane Glycoproteins; D008854:Microscopy, Electron; D009243:NAD / Q000494:pharmacology; D010053:Ovary; D010084:Oxidation-Reduction; D011104:Polyisoprenyl Phosphate Oligosaccharides / Q000096:biosynthesis; D014415:Tunicamycin / Q000494:pharmacology; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000378:metabolism*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011104:Polyisoprenyl Phosphate Oligosaccharides; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D009243:NAD; D014415:Tunicamycin; D000117:Acetylglucosamine