Baker R R, Chang H Y
Biochem Cell Biol. 1986 Jan;64(1):1-7. doi: 10.1139/o86-001.
A neuronal nuclear fraction (N1) was isolated from cerebral cortices of 15-day-old rabbits. Samples of N1 were incubated with a radioactive fatty acid ([3H]arachidonate or [14C]oleate), acylation cofactors, and 1-acyl-sn-glycero-3-phosphoinositol (1-acyl-GPI) or 1-acyl-sn-glycero-3-phosphocholine (1-acyl-GPC). In competition studies, both radioactive fatty acids were incubated with one lysophospholipid or the two lysophospholipids were incubated with one radioactive fatty acid. Using [3H] arachidonate and one lysophosphoglyceride, a maximal rate of incorporation into phosphatidylinositol (PI) was found at a relatively low concentration of 1-acyl-GPI (10 microM), while increasing rates of incorporation into phosphatidylcholine (PC) were seen with increasing concentrations of 1-acyl-GPC (to 65 microM). At low concentrations of lysophosphoglyceride (less than or equal to 25 microM) the rate of arachidonate incorporation into PI greatly exceeded rates of arachidonate incorporation into PC. This higher rate of arachidonate incorporation into PI was also seen in incubations where both lysophospholipids were present. For oleate, greater rates of incorporation into PC were found in comparison with rates of labelling of PI in assays using relatively high concentrations of one or both lysophospholipids. When comparing arachidonate and oleate, in assays with one or both fatty acids, the polyunsaturate showed at least threefold higher rates of incorporation into PI. For PC labelling higher rates of arachidonate incorporation were evident at the higher concentrations of 1-acyl-GPC and the superiority over oleate was not as marked as that seen in PI labelling.
从15日龄兔的大脑皮层中分离出神经元核组分(N1)。将N1样品与放射性脂肪酸([3H]花生四烯酸或[14C]油酸)、酰化辅因子以及1-酰基-sn-甘油-3-磷酸肌醇(1-酰基-GPI)或1-酰基-sn-甘油-3-磷酸胆碱(1-酰基-GPC)一起孵育。在竞争研究中,将两种放射性脂肪酸与一种溶血磷脂一起孵育,或者将两种溶血磷脂与一种放射性脂肪酸一起孵育。使用[3H]花生四烯酸和一种溶血甘油酯,发现在相对较低浓度的1-酰基-GPI(10 microM)时,花生四烯酸掺入磷脂酰肌醇(PI)的速率最高,而随着1-酰基-GPC浓度增加(至65 microM),掺入磷脂酰胆碱(PC)的速率也增加。在低浓度的溶血甘油酯(小于或等于25 microM)下,花生四烯酸掺入PI的速率大大超过其掺入PC的速率。在两种溶血磷脂都存在的孵育实验中也观察到花生四烯酸掺入PI的速率较高。对于油酸,在使用相对较高浓度的一种或两种溶血磷脂的实验中,发现其掺入PC的速率高于标记PI的速率。当比较花生四烯酸和油酸时,在使用一种或两种脂肪酸的实验中,多不饱和脂肪酸掺入PI的速率至少高3倍。对于PC标记,在较高浓度的1-酰基-GPC下,花生四烯酸的掺入速率更高,并且其相对于油酸的优势不如在PI标记中那么明显。