Biosynthesis of phosphatidylcholine from serum phospholipids in Chinese hamster ovary cells deprived of choline.
We report that Chinese hamster ovary cells assemble membrane phospholipids from choline-linked lipid present in fetal calf serum. This was examined by testing the ability of various serum preparations to satisfy the choline requirement of the cells. Chinese hamster ovary cells divided in growth medium containing lipoprotein-deficient serum and approximately 8 microM lysolecithin. Identical results were obtained in growth medium supplemented with solvent-extracted (delipidated) serum reconstituted with purified egg lysolecithin and the uptake of lipid was inhibited by the addition of bovine serum albumin. Analysis of the phospholipid composition of cells incubated with 32Pi and egg lysolecithin in place of choline revealed that approximately 30% of the phosphorus moieties of the cellular phospholipids were derived from the added lipid, while in the presence of choline less than 10% arose in this fashion. Choline starvation enhanced the formation of lecithin from [32P]lysolecithin without affecting phospholipid turnover and labeled lecithin was converted to other phospholipids, especially sphingomyelin and phosphatidylserine. Unlike endogenous serum lysolecithin, lipoproteins obtained from human and fetal calf sera failed to satisfy the choline requirement of Chinese hamster ovary cells, even though 95% of the lipoprotein phospholipid was phosphatidylcholine and sphingomyelin. Together, these results demonstrate that animal cells can derive all of the choline required for membrane phospholipid synthesis from serum lysolecithin and that its conversion to lecithin within the cell is regulated by the availability of choline. In contrast, serum lipoproteins do not normally serve as a major source of choline moieties.
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002794:Choline / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D005260:Female; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood; D010053:Ovary; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010743:Phospholipids / Q000097:blood / Q000378:metabolism*
D003470:Culture Media; D008077:Lipoproteins, LDL; D010713:Phosphatidylcholines; D010743:Phospholipids; D002794:Choline