Latency of the peroxisomal enzyme acyl-CoA:dihydroxyacetonephosphate acyltransferase in digitonin-permeabilized fibroblasts: the effect of ATP and ATPase inhibitors.
We have studied the activity of acyl-CoA:dihydroxyacetonephosphate acyltransferase (DHAP-AT) in fibroblasts treated with low concentrations of digitonin so that the cytoplasmic compartment was freely accessible to the substrates of DHAP-AT while intracellular membranes remained intact. DHAP-AT activity exhibited 70% latency under these conditions. This latency could be overcome by addition of ATP, resulting in a four-fold stimulation of DHAP-AT activity. Virtually no stimulatory effect of ATP on DHAP-AT activity was observed in sonicated fibroblasts or when a non-hydrolyzable ATP analogue was used. Furthermore the stimulatory effect of ATP was prevented in part by DCCD. N-ethylmaleimide and high concentrations of oligomycin; bafilomycin had no effect. This pattern of inhibitor sensitivity is similar to that of the ATPase activity in peroxisomal fractions from rat liver. We conclude that peroxisomes in situ exhibit structure linked latency and that ATP is required for the transport of at least one of the substrates of DHAP-AT.
Wolvetang
E J
EJ
Department of Pediatrics, Academic Medical Centre, Amsterdam, The Netherlands.
Tager
J M
JM
Wanders
R J
RJ
D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D004072:Digitonin / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008830:Microbodies / Q000201:enzymology*; D013010:Sonication
C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D000217:Acyltransferases; C020051:glycerone-phosphate O-acyltransferase; D000251:Adenosine Triphosphatases; D004072:Digitonin