Schroeder F, Doi O
Biochim Biophys Acta. 1978 Oct 25;531(1):32-43. doi: 10.1016/0005-2760(78)90179-0.
Incorporation and loss of membrane lipids were measured in murine fibroblasts grown in suspension culture with [2-14C]acetate in a chemically defined medium devoid of serum or lipid. Greater than 98 and 94% of the 14C present in the phospholipids and neutral glycerides respectively was found in the fatty acyl groups. During a 3-day growth period LM fibroblasts lost 65% of their 14C-labelled acyl groups from the phospholipids and 36% from the neutral lipids. Selective retention of 14C in individual isolated phospholipid and neutral lipid species occurred only in phosphatidylethanolamine. Retention of acyl groups in the phospholipids or neutral glycerides was not a property of LM suspension cultured fibroblasts. Supplementation of the LM cells with choline analogues, such as N,N'-dimethylethanolamine, N-monomethylethanolamine, or ethanolamine instead of choline, decreased the loss of 14C-labelled fatty acids from phospholipid and neutral lipid. Ethanolamine supplementation resulted in increased loss of 14C from desmesterol as well as decreased incorporation of 14C into desmosterol. Thus, polar head group manipulation affected acyl group composition, acyl group retention, and sterol metabolism in suspension cultured LM cells.
在无血清或脂质的化学限定培养基中,用[2-¹⁴C]乙酸盐培养悬浮的小鼠成纤维细胞,测量膜脂的掺入和损失。分别在磷脂和中性甘油酯中的¹⁴C中,超过98%和94%存在于脂肪酰基中。在3天的生长期间,LM成纤维细胞从磷脂中损失了65%的¹⁴C标记的酰基,从中性脂质中损失了36%。¹⁴C在单个分离的磷脂和中性脂质种类中的选择性保留仅发生在磷脂酰乙醇胺中。酰基在磷脂或中性甘油酯中的保留不是LM悬浮培养成纤维细胞的特性。用胆碱类似物,如N,N'-二甲基乙醇胺、N-单甲基乙醇胺或乙醇胺代替胆碱补充LM细胞,可减少¹⁴C标记的脂肪酸从磷脂和中性脂质中的损失。补充乙醇胺导致¹⁴C从羊毛甾醇中的损失增加,以及¹⁴C掺入羊毛甾醇的减少。因此,极性头部基团的操作影响了悬浮培养的LM细胞中的酰基组成、酰基保留和甾醇代谢。