Morash S C, Cook H W, Spence M W
Atlantic Research Centre for Mental Retardation, Dalhousie University, Halifax, Canada.
Biochim Biophys Acta. 1988 Jul 22;961(2):194-202. doi: 10.1016/0005-2760(88)90114-2.
The catabolism of phosphatidylcholine (PtdCho) has been studied in cultured murine neuroblastoma (N1E-115), C6 glioma, rat brain primary glia, and human fibroblast cells. Cells were pulse labelled for 96 h with [methyl-3H]choline followed by a chase for up to 24 h in medium containing 4 mM choline. Measurement of the radioactivity and mass of choline-containing compounds in these cells indicated that the major degradative pathway is PtdCho----lysophosphatidylcholine (lysoPtdCho)----glycerophosphocholine (GroPCho)----choline. At all times during the chase, PtdCho, sphingomyelin and lysoPtdCho comprised 72-92% of the cell-associated radioactivity; the remaining 10-30% was water-soluble and was chiefly GroPCho (30-80%) in all cell lines. In fibroblasts, however, phosphocholine (PCho) was also a major labelled water-soluble component (33-54%). The specific activity of GroPCho closely parallelled that of PtdCho in fibroblasts, but decreased faster than PtdCho in C6 and N1E-115 cells. We postulate that this may be due to distinct pools of PtdCho in the cell with differing rates of turnover. The changes in specific activity of PCho suggest that the major portion is formed by synthesis rather than as a degradative product. However, the inability to reduce the specific activity of this fraction to that of the intracellular choline suggests that a portion may be derived from either PtdCho or GroPCho.
已在培养的小鼠神经母细胞瘤(N1E - 115)、C6胶质瘤、大鼠脑原代神经胶质细胞和人成纤维细胞中研究了磷脂酰胆碱(PtdCho)的分解代谢。用[甲基 - ³H]胆碱对细胞进行96小时的脉冲标记,然后在含有4 mM胆碱的培养基中追踪长达24小时。对这些细胞中含胆碱化合物的放射性和质量的测量表明,主要的降解途径是PtdCho→溶血磷脂酰胆碱(lysoPtdCho)→甘油磷酸胆碱(GroPCho)→胆碱。在追踪过程中的所有时间,PtdCho、鞘磷脂和lysoPtdCho占细胞相关放射性的72 - 92%;其余10 - 30%是水溶性的,在所有细胞系中主要是GroPCho(30 - 80%)。然而,在成纤维细胞中,磷酸胆碱(PCho)也是一种主要的标记水溶性成分(33 - 54%)。在成纤维细胞中,GroPCho的比活性与PtdCho的比活性密切平行,但在C6和N1E - 115细胞中比活性下降得比PtdCho更快。我们推测这可能是由于细胞中具有不同周转率的不同PtdCho池所致。PCho比活性的变化表明,其主要部分是通过合成形成的,而不是作为降解产物。然而,无法将该部分的比活性降低到细胞内胆碱的比活性水平,这表明一部分可能来自PtdCho或GroPCho。