Nelsen V M, Mackenzie C G, Resis O K, Mackenzie J B, Mortiz E
Biochim Biophys Acta. 1975 May 22;388(2):188-97. doi: 10.1016/0005-2760(75)90123-x.
Rabbit liver cells, in which fatty acid synthesis was suppressed by the rabbit serum component of the medium, were grown through 8- to 120-fold increases in cell numbers and mass of cell lipid in the presence of [U-14 C]-glucose. Triacylglycerols, phosphatidylcholine, and phosphatidylethanolamine were isolated from the total cell lipid and deacylated. Carbons 1 and 3 of the glycerol from the triacylglycerols and the no. 1 glycerol carbons of the two deacylated phospholids were oxidized by periodate and isolated as the dimedon derivative of formaldehyde. The specific activities of the glycerol carbons indicated that 58, 44, and 37 percent of the glycerol of the triacylglycerols. phosphatidylcholine, and phosphatidylethanolamine, respectively, were derived from the glucose of the medium. An additional 8 percent and 1-2 percent of the glycerol of each lipid was derived, respectively, from [U-14 C] glycerol and U14C-labeled amino acids added to the medium. In agreement with an experiment with albumin-bound [9,10- minus 3H]-oleic acid, and with smilar earlier experiments, it appears likely that appriacylglycerols originated from serum lipoproteins, or their partial hydrolysis products. An appreciable part of the ethanolamine of the cells' phosphatidylethanolamine originated from exogenous U- minus 14 C-labeled amino acids. Phosphatidyl-ethanolamine, however, was not a primary source of phosphatidylcholine. Labeling of the fatty acids of triacylglycerols and phospholipids by radioactive glucose, glycerol and amino acids was negligible.
兔肝细胞在培养基中的兔血清成分抑制脂肪酸合成的情况下,在[U-14C] -葡萄糖存在下生长,细胞数量增加了8至120倍,细胞脂质质量也增加。从总细胞脂质中分离出三酰甘油、磷脂酰胆碱和磷脂酰乙醇胺并进行脱酰基处理。三酰甘油中甘油的碳1和碳3以及两种脱酰基磷脂的1号甘油碳被高碘酸盐氧化,并作为甲醛的二甲基苯腙衍生物分离出来。甘油碳的比活性表明,三酰甘油、磷脂酰胆碱和磷脂酰乙醇胺中分别有58%、44%和37%的甘油来自培养基中的葡萄糖。每种脂质另外分别有8%和1 - 2%的甘油来自添加到培养基中的[U-14C]甘油和U14C标记的氨基酸。与用白蛋白结合的[9,10-3H] -油酸进行的实验以及早期类似实验一致,三酰甘油似乎起源于血清脂蛋白或其部分水解产物。细胞磷脂酰乙醇胺中相当一部分乙醇胺起源于外源性U-14C标记的氨基酸。然而,磷脂酰乙醇胺不是磷脂酰胆碱的主要来源。放射性葡萄糖、甘油和氨基酸对三酰甘油和磷脂脂肪酸的标记可以忽略不计。