Kuwae T, Schmid P C, Schmid H H
Faculty of Pharmaceutical Sciences, Josai University, Saitama, Japan.
Biochim Biophys Acta. 1997 Jan 7;1344(1):74-86. doi: 10.1016/s0005-2760(96)00135-x.
Glycerophospholipid biosynthesis by the de novo pathway was assessed in mouse peritoneal macrophages by pulse-labeling with [U-14C]glycerol. Phosphatidylcholine (PC), which amounts to about 35% of total cellular phospholipids, exhibited the highest rate of glycerol uptake, followed by phosphatidylinositol (PI) and phosphatidylethanolamine (PE). Remodeling of PC molecular species by deacylation/reacylation was established by determining the redistribution of glycerol label over 2 h after a 1 h pulse of [U-14C]glycerol and by determining incorporation of 18O from H2(18)O-containing media. These data suggest that stearic and arachidonic acid enter PC primarily by the remodeling pathway but that small amounts of highly unsaturated molecular species, including 1,2-diarachidonoyl PC, are rapidly synthesized de novo, and subsequently remodeled or degraded. Treatment of the cells with the ionophore A23187 resulted in the selective enhancement of arachidonate turnover in PC, PI and neutral lipid, as well as enhanced de novo PI synthesis. [U-14C]Glycerol labeling experiments suggest that arachidonic acid liberated by Ca(2+)-dependent phospholipase A2 activity is also reacylated in part through de novo glycerolipid biosynthesis, leading to the formation and remodeling of 1,2-diarachidonoyl PC and other highly polyunsaturated molecular species.
通过用[U-14C]甘油进行脉冲标记,在小鼠腹腔巨噬细胞中评估了从头合成途径的甘油磷脂生物合成。磷脂酰胆碱(PC)约占细胞总磷脂的35%,其甘油摄取率最高,其次是磷脂酰肌醇(PI)和磷脂酰乙醇胺(PE)。通过在[U-14C]甘油脉冲1小时后测定2小时内甘油标记的重新分布,以及通过测定含H2(18)O培养基中18O的掺入,确定了通过脱酰基/再酰化对PC分子种类的重塑。这些数据表明,硬脂酸和花生四烯酸主要通过重塑途径进入PC,但少量高度不饱和分子种类,包括1,2-二花生四烯酰基PC,是迅速从头合成的,随后进行重塑或降解。用离子载体A23187处理细胞导致PC、PI和中性脂质中花生四烯酸周转的选择性增强,以及从头PI合成增强。[U-14C]甘油标记实验表明,由钙依赖性磷脂酶A2活性释放的花生四烯酸也部分通过从头甘油脂质生物合成重新酰化,导致1,2-二花生四烯酰基PC和其他高度多不饱和分子种类的形成和重塑。