Different contribution of phospholipid and triacylglycerol metabolism to esterification of free intracellular arachidonate: a study on SK-N-BE(2) human neuroblastoma cells.
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When SK-N-BE(2) human neuroblastoma cells were exposed for 1h to growth medium supplemented with [14C]arachidonic acid (AA) at final concentrations ranging from 1 microM to 100 microM, an amount of this fatty acid was uptaken ranging form a 2% to a 120% of that present in cells at steady state. As more [14C]AA was uptaken by cells, a larger fraction was progressively incorporated into triacylglycerols (TAG) in comparison to phospholipids (PL), with minor amounts remaining in a free form. By gas chromatographic analysis it was estimated that TAG from cells grown in ordinary medium contained about 2 nmoles AA per mg protein, but, after 1 h exposure to medium supplemented with 100 microM AA (label-free) this value rose to about 28 nmoles/mg protein; furthermore, as estimated on the basis of total fatty acid content, TAG mass was increased by a 16%. Cell exposure to medium enriched with 100 microM AA did not cause PL mass changes, whereas AA content was significantly increased only in phosphatidylcholine. Medium enrichment with 100 microM AA dramatically enhanced [3H]glycerol incorporation into TAG, as assessed after 1 h cell pulse, with minor but significant changes observed also for phosphatidylinositol and phosphatidylethanolamine, but not for phosphatidylcholine. In the light of these data, the contribution of PL and TAG to the removal of free intracellular AA is discussed.