Stoffel W, Bister K
Biochemistry. 1975 Jul;14(13):2841-7. doi: 10.1021/bi00684a008.
13-C nuclear magnetic resonance (NMR) studies are described regarding the lipid organization in the envelope of the vesicular stomatitis virion. The fatty acid chains (oleic acid) and the choline moiety of the 3-sn-phosphatidylcholine and spingomyelin have been labeled specifically with 13-C by growing the virions in prelabeled host cells (BHK 21 cells). The results suggest that 130C NMR spectroscopy is a very feasible method for the study of natural membranes provided the isotope is highly enriched in specific positions and incorporated biochemically. Spin-lattice relaxation (T1) measurements of particular C atoms have been carried out with whole virions, with virions deprived of their surface projections by trypsinization but unaltered in their shape and size, and with liposomes prepared from the total lipid mixture of the envelope in order to get insight into the molecular structure of this model membrane. The mobility of the central part of 11-13-C-labeled oleic acid incorporated into the ester and amide lipids and the choline group of 3-sn-phosphatidylcholine and sphingomyelin is very restricted as indicated by their short T1 times. It is concluded from the data presented here that the high cholesterol content (cholesterol/P: 0.7) of the envelope lipid phase is responsible for the rather rigidly packed envelope structure. The mode and extent of the interactions between lipids and glycoprotein surface projections are subjects for further study.
描述了关于水泡性口炎病毒粒子包膜中脂质组织的13C核磁共振(NMR)研究。通过在预先标记的宿主细胞(BHK 21细胞)中培养病毒粒子,3 - sn - 磷脂酰胆碱和鞘磷脂的脂肪酸链(油酸)以及胆碱部分已被13C特异性标记。结果表明,只要该同位素在特定位置高度富集并通过生化方式掺入,13C NMR光谱法是研究天然膜的一种非常可行的方法。已对完整病毒粒子、经胰蛋白酶处理去除其表面突起但形状和大小未改变的病毒粒子以及由包膜的总脂质混合物制备的脂质体进行了特定碳原子的自旋 - 晶格弛豫(T1)测量,以便深入了解该模型膜的分子结构。掺入酯类和酰胺类脂质中的11 - 13C标记油酸的中心部分以及3 - sn - 磷脂酰胆碱和鞘磷脂的胆碱基团的流动性非常受限,这由它们较短的T1时间表明。从这里给出的数据可以得出结论,包膜脂质相的高胆固醇含量(胆固醇/磷脂:0.7)是包膜结构紧密堆积的原因。脂质与糖蛋白表面突起之间相互作用的方式和程度是有待进一步研究的课题。