Gsalpha-mediated regulation of the carnitine carrier in S49 lymphoma cells.
Carnitine is essential for mitochondrial oxidation of long-chain fatty acids. Peripheral cells rely on plasma transport of carnitine which is taken up by an active mechanism in the plasma membrane. This project investigated the plasma membrane bound carnitine carrier in cultured S49 lymphoma cells. We investigated wild-type cells and two mutant cells lines showing deficient activity of adenylyl cyclase, cyc- lacking and H21a containing a deficient Gsalpha. Plasma membranes derived from cyc- cells displayed six times more carnitine binding sites and a 1.35 times faster uptake rate than plasma membranes from wild-type cells. In vitro mixing of plasma membranes from cyc- and wild-type cells transferred a factor reducing the number of expected carnitine binding sites by about 30%. Cyclic AMP could not substitute for wild-type membranes as the inhibitor of carnitine binding to plasma membranes derived from cyc- cells. Cholera toxin induced ADP-ribosylation of Gsalpha causing activation of Gsalpha present in wild-type but not in cyc- cells, further reducing carnitine uptake and carnitine binding to plasma membranes. Our findings thus supported the notion that Gsalpha by a mechanism not involving cyclic AMP inhibited cellular uptake of carnitine by reducing the number of available carnitine binding sites in plasma membranes.
Department of Allergology, Sahlgren's University Hospital, S-413 45 Göteborg, Sweden.
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Gustafson
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Ransnäs
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000172:deficiency / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001692:Biological Transport; D002331:Carnitine / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002772:Cholera Toxin / Q000494:pharmacology; D003470:Culture Media; D000242:Cyclic AMP / Q000494:pharmacology; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000502:physiology*; D007700:Kinetics; D008223:Lymphoma / Q000378:metabolism*; D051379:Mice; D014407:Tumor Cells, Cultured
D003470:Culture Media; D000246:Adenosine Diphosphate Ribose; D002772:Cholera Toxin; D000242:Cyclic AMP; D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases; D002331:Carnitine