Schneider A, Eichenberger W, Seebeck T
Institut für Allgemeine Mikrobiologie, Universität Bern, Switzerland.
J Biol Chem. 1988 May 15;263(14):6472-5.
Covalently linked fatty acids are increasingly recognized as an important type of post-translational protein modification. Many of such acylated proteins are found associated with cellular membranes. The membrane skeleton of the parasitic hemoflagellate Trypanosoma brucei consists of a regular array of microtubules which are tightly bound to the overlying cell membrane. A microtubule-binding protein (p41) has been identified within this structure which carries covalently bound fatty acid. The fatty acid linkage is sensitive to hydroxylamine treatment. After chemical transesterification, the released radioactivity co-migrates with fatty acid methyl esters in thin-layer chromatograms, establishing that the fatty acid was covalently bound to p41 via an ester (thioester) linkage. Upon detergent extraction of trypanosomes, p41 remains tightly bound to the cytoskeleton as long as Ca2+ ions are present. It can selectively be released from this structure by the addition of excess EGTA. Conversely, p41 binds to isolated cytoskeletons and to purified microtubules in vitro, the reaction again being entirely Ca2+-dependent.
共价连接的脂肪酸越来越被认为是一种重要的翻译后蛋白质修饰类型。许多这类酰化蛋白被发现与细胞膜相关。寄生性血鞭毛虫布氏锥虫的膜骨架由规则排列的微管组成,这些微管紧密结合在覆盖其上的细胞膜上。在这个结构中已鉴定出一种携带共价连接脂肪酸的微管结合蛋白(p41)。脂肪酸连接对羟胺处理敏感。化学酯交换后,释放的放射性物质在薄层色谱图中与脂肪酸甲酯共同迁移,这表明脂肪酸通过酯(硫酯)键与p41共价结合。在用去污剂提取锥虫后,只要存在Ca2+离子,p41就会紧密结合在细胞骨架上。通过添加过量的乙二醇双四乙酸(EGTA),它可以从这个结构中选择性地释放出来。相反,p41在体外与分离的细胞骨架和纯化的微管结合,该反应同样完全依赖于Ca2+。