The molecular selectivity of phospholipase D in HL60 granulocytes.
The molecular selectivity of PLD in PMA-stimulated HL60 granulocytes was determined by HPLC analysis of [3H]butanol incorporation into phosphatidyl[3H]butanol (Ptd[3H]But) molecular species. Comparison with phospholipid compositions confirmed that PLD acted primarily on phosphatidylcholine (PtdCho). Apparent enzyme selectivity was suggested by negligible formation of PB16:0/16:0 and preferential synthesis of Ptd[3H]But species containing sn-1 18:0. Culture with exogenous 18:2n-6 or 20:4n-6 readily modified both PtdCho and Ptd[3H]But compositions, and accentuated the apparent selectivity of stimulated PLD for sn-1 18:0 species of PtdCho. Such modifications to PLD-based signalling mechanisms may contribute to the modulatory effects of altered dietary lipid intakes on cellular functions.
Heung
Y M
YM
University of Southampton, Southampton General Hospital, UK.
Postle
A D
AD
D000440:Butanols / Q000378:metabolism; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D020404:Glycerophospholipids; D006098:Granulocytes / Q000201:enzymology*; D006801:Humans; D010712:Phosphatidic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014316:Tritium
D000440:Butanols; D020404:Glycerophospholipids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; C068564:phosphatidylbutanol; D014316:Tritium; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate