Fatty alcohol metabolism in cultured human fibroblasts. Evidence for a fatty alcohol cycle.
Intact cultured human fibroblasts reduced [1-14C]palmitate to radioactive hexadecanol in a concentration-dependent manner. In the presence of 30 microM radioactive palmitate, cellular levels of labeled hexadecanol increased over time and reached a steady state corresponding to at least 0.1% of cell-associated radioactive palmitate. These levels of [14C]hexadecanol were increased up to 10-fold when exogenous nonradioactive hexadecanol was present, suggesting that radioactive hexadecanol was actively metabolized. Cells incubated in fatty acid-free medium with [1-14C]hexadecanol rapidly oxidized it to palmitic acid; less than 2% of the hexadecanol taken up by the cells was incorporated into the ether linkage of phosphatidylethanolamine, and no incorporation into wax esters was detected. Double-label experiments involving incubation of intact fibroblast with [3H]palmitate and [14C]hexadecanol demonstrated simultaneous synthesis of hexadecanol from palmitate and oxidation of hexadecanol to palmitate. Addition of exogenous palmitate to the medium of intact cells inhibited the oxidation of hexadecanol to fatty acid in a concentration-dependent fashion. This was associated with an increase in the fibroblast content of hexadecanol and loss of hexadecanol into the medium. Activity of fatty alcohol:NAD+ oxidoreductase, which catalyzes the oxidation of hexadecanol to palmitic acid, was inhibited by palmitoyl-CoA and NADH, but not by palmitic acid. These results are consistent with the presence of a "fatty alcohol cycle" in which hexadecanol is synthesized from palmitate via acyl-CoA and simultaneously oxidized back to free fatty acid. Fatty acyl-CoA, which is the primary substrate for fatty alcohol synthesis, may also regulate the intracellular level of fatty alcohol by inhibiting its oxidation.
Rizzo
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WB
Department of Pediatrics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298.
Craft
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DA
Dammann
A L
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Phillips
M W
MW
D000214:Acyl Coenzyme A / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D005233:Fatty Alcohols / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D009243:NAD / Q000378:metabolism; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D010171:Palmitoyl Coenzyme A / Q000494:pharmacology
D000214:Acyl Coenzyme A; D005233:Fatty Alcohols; D010169:Palmitic Acids; D009243:NAD; D010171:Palmitoyl Coenzyme A; D019308:Palmitic Acid; C005031:cetyl alcohol; D000429:Alcohol Oxidoreductases; C065402:long-chain-alcohol dehydrogenase