Two distinct members of the ADP-ribosylation factor family of GTP-binding proteins regulate cell-free intra-Golgi transport.
We have used an intra-Golgi transport assay to identify GTP-binding proteins involved in regulation of protein traffic. Two soluble proteins of 20 kd were purified by their ability to mediate GTP gamma S-dependent inhibition of transport. These GTP-dependent Golgi binding factors, or GGBFs, exhibit a 3-fold difference in activity and are differentiated by their hydrophobicity, isoelectric points, and apparent size. Removal of 80% of GGBFs from cytosol abolishes GTP gamma S sensitivity but does not affect inhibition by aluminum fluoride. We demonstrate that GGBFs are members of the ADP-ribosylation factor (ARF) family. Recombinant ARF1 exhibits GGBF activity and myristoylation is required. The distinct biochemical properties of GGBFs indicate that members of the ARF family may have related but distinct functions in intracellular transport.
Taylor
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Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Kahn
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Melançon
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D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors; D000535:Aluminum / Q000494:pharmacology; D017607:Aluminum Compounds; D001692:Biological Transport / Q000187:drug effects; D020126:Brefeldin A; D002474:Cell-Free System; D003517:Cyclopentanes / Q000494:pharmacology; D005459:Fluorides / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D014764:Viral Proteins / Q000378:metabolism*
D017607:Aluminum Compounds; D003517:Cyclopentanes; D014764:Viral Proteins; D020126:Brefeldin A; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000535:Aluminum; D019204:GTP-Binding Proteins; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors; D005459:Fluorides; C032311:aluminum fluoride