Phospholipase D is present on Golgi-enriched membranes and its activation by ADP ribosylation factor is sensitive to brefeldin A.
ADP ribosylation factor (ARF) is a small guanosine triphosphate (GTP)-binding protein that regulates the binding of coat proteins to membranes and is required for several stages of vesicular transport. ARF also stimulates phospholipase D (PLD) activity, which can alter the lipid content of membranes by conversion of phospholipids into phosphatidic acid. Abundant PLD activity was found in Golgi-enriched membranes from several cell lines. Golgi PLD activity was greatly stimulated by ARF and GTP analogs and this stimulation could be inhibited by brefeldin A (BFA), a drug that blocks binding of ARF to Golgi membranes. Furthermore, in Golgi membranes from BFA-resistant PtK1 cells, basal PLD activity was high and not stimulated by exogenous ARF or GTP analogs. Thus, ARF activates PLD on the Golgi complex, suggesting a possible link between transport events and the underlying architecture of the lipid bilayer.
Ktistakis
N T
NT
Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas 72935-9038, USA.
Brown
H A
HA
Sternweis
P C
PC
Roth
M G
MG
D020727:ADP-Ribosylation Factors; D000818:Animals; D020126:Brefeldin A; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism; D002458:Cell Fractionation; D002460:Cell Line; D006224:Cricetinae; D003517:Cyclopentanes / Q000494:pharmacology*; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology* / Q000648:ultrastructure; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000201:enzymology* / Q000648:ultrastructure; D007700:Kinetics; D009975:Orthomyxoviridae / Q000235:genetics; D010739:Phospholipase D / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*
D002352:Carrier Proteins; D003517:Cyclopentanes; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D010739:Phospholipase D; D019204:GTP-Binding Proteins; D020727:ADP-Ribosylation Factors