Rochester C P, Bishop D G
Arch Biochem Biophys. 1984 Jul;232(1):249-58. doi: 10.1016/0003-9861(84)90541-1.
The incorporation of oleate from oleoyl-CoA into lipids by microsomes from developing sunflower (Helianthus annuus L.) seeds has been investigated. Oleate was incorporated mainly into position 2 of phosphatidylcholine or released as free fatty acid. The addition of exogenous 1-acyl-lysophosphatidylcholine increased the incorporation of oleate into position 2 of phosphatidylcholine and decreased the release of free oleate. In the absence of exogenous lysophosphatidylcholine, the incorporation of oleate into phosphatidylcholine was limited by the amount of endogenous acceptor present. DH-990, an inhibitor of acyl-CoA:lysophosphatidylcholine acyltransferase, almost completely inhibited the incorporation of oleate from oleoyl-CoA into phosphatidylcholine at a concentration of 2.5 mM. These results indicate that the incorporation of oleate from oleoyl-CoA into microsomal phosphatidylcholine occurs mainly by the acylation of a 1-acyl-lysophosphatidylcholine acceptor rather than by acyl exchange between oleoyl-CoA and phosphatidylcholine. While the incorporation of oleoyl-CoA was completed within 2 to 5 min, exogenous 1-acyl-lysophosphatidylcholine was incorporated into phosphatidylcholine for up to 30 min. Addition of oleoyl-CoA resulted in an increase in both the rate and magnitude of lysophosphatidylcholine incorporation, which could not be accounted for by a stoichiometric reaction between the two substrates. Evidence is provided that free CoA had an independent stimulatory effect on the incorporation of lysophosphatidylcholine. The implications of this finding are discussed.
对发育中的向日葵(Helianthus annuus L.)种子微粒体将油酰辅酶A中的油酸酯掺入脂质的过程进行了研究。油酸酯主要掺入磷脂酰胆碱的2位或作为游离脂肪酸释放。添加外源性1-酰基溶血磷脂酰胆碱可增加油酸酯掺入磷脂酰胆碱2位的量,并减少游离油酸的释放。在没有外源性溶血磷脂酰胆碱的情况下,油酸酯掺入磷脂酰胆碱受到内源性受体数量的限制。酰基辅酶A:溶血磷脂酰胆碱酰基转移酶抑制剂DH-990在浓度为2.5 mM时几乎完全抑制了油酰辅酶A中的油酸酯掺入磷脂酰胆碱。这些结果表明,油酰辅酶A中的油酸酯掺入微粒体磷脂酰胆碱主要是通过1-酰基溶血磷脂酰胆碱受体的酰化作用,而不是通过油酰辅酶A与磷脂酰胆碱之间的酰基交换。虽然油酰辅酶A的掺入在2至5分钟内完成,但外源性1-酰基溶血磷脂酰胆碱掺入磷脂酰胆碱的时间长达30分钟。添加油酰辅酶A导致溶血磷脂酰胆碱掺入的速率和幅度均增加,这不能用两种底物之间的化学计量反应来解释。有证据表明游离辅酶A对溶血磷脂酰胆碱的掺入有独立的刺激作用。讨论了这一发现的意义。