Rampini C, Dubois C
Département de Biochimie, Biologie Moléculaire et Biologie Cellulaire, Faculté de Médecine Saint-Antoine, Paris, France.
Biochim Biophys Acta. 1995 Jun 27;1257(1):75-80. doi: 10.1016/0005-2760(95)00056-i.
Phospholipid synthesis was investigated in concanavalin A-activated human peripheral lymphocytes up until 72 h following cell activation, i.e., during the G1 and S phases of the cell cycle. Using [32P]phosphate pulse experiments (5 h), striking differences were observed between phosphatidylethanolamine (PE) and phosphatidylcholine (PC) synthesis. Both the incorporation of [32P]phosphate into PE and the PE/PC incorporation ratio were greatly enhanced, 16-fold and 8-fold, respectively, after 48 h of incubation with the mitogen. This increase in PE synthesis was still observed when cell entry into the S phase was inhibited by an excess of concanavalin A; thereby it must be related to the late stages of the G1 phase. The stimulation of the incorporation into PE was the same for both [14C]ethanolamine and [32P]phosphate, therefore suggesting the involvement of the phosphoethanolamine synthesis pathway. Kinetics of continuous incorporation of [32P]phosphate into PE and PC indicated that the PE/PC net synthesis ratio was enhanced in activated cells, which corresponds to PE enrichment in lymphocyte membranes. The stimulation of PE synthesis in late G1 may be of importance for cell progression through the cell cycle by changing the membrane physical properties. Furthermore, it may serve as a test for checking lymphocyte reactivity to mitogens.
在刀豆球蛋白A激活的人外周血淋巴细胞中,对磷脂合成进行了研究,观察时间长达细胞激活后的72小时,即细胞周期的G1期和S期。通过[32P]磷酸盐脉冲实验(5小时),观察到磷脂酰乙醇胺(PE)和磷脂酰胆碱(PC)合成之间存在显著差异。在用有丝分裂原孵育48小时后,[32P]磷酸盐掺入PE的量以及PE/PC掺入比均大幅增加,分别增加了16倍和8倍。当过量的刀豆球蛋白A抑制细胞进入S期时,仍观察到PE合成的增加;因此,这一定与G1期的后期阶段有关。[14C]乙醇胺和[32P]磷酸盐掺入PE的刺激作用相同,因此表明磷酸乙醇胺合成途径参与其中。[32P]磷酸盐持续掺入PE和PC的动力学表明,激活细胞中PE/PC的净合成比增加,这与淋巴细胞膜中PE的富集相对应。G1期后期PE合成的刺激作用可能通过改变膜的物理性质对细胞通过细胞周期的进程具有重要意义。此外,它还可以作为检测淋巴细胞对有丝分裂原反应性的一种测试。