放射性示踪法研究结合希土的四膜虫膜蛋白——I.铈、镱、铥膜蛋白提取与分子量测定
A STUDY OF RARE EARTH BINDING MEMBRANE PROTEIN OF TETRAHYMENA PYRIFORMIS WITH RADIOTRACEP. I. EXTRACTION OF Ce, Yb, Tm BINDING MEMBRANE PROTEIN AND ITS MOL. WT. DETERMINATION
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摘要: 四膜虫细胞在分别加入了示踪量放射性希土离子~(141)Ce(Ⅲ),~(169)Yb(Ⅲ),~(170)Tm(Ⅲ)的培养液中生长,实验确定65%的放射性集中在细胞膜上。使用生物化学离心分离程序,除去细胞纤毛和线粒体等,得到了细胞膜。膜蛋白是用去污剂2%去氧胆汁酸钠处理细胞膜提取的。提取液用葡聚糖G-100凝胶层析柱分离出膜蛋白的组分,层析图上蛋白洗脱峰与放射性峰吻合,表明所得的是一种与希土离子较牢固地结合的蛋白质。进一步用SDS-聚丙烯酰胺凝胶电泳法,将层析柱的峰位膜蛋白物质与标准系列蛋白质同时电泳,测得含希土膜蛋白亚基的分子量为54000。Abstract: Tetrahymena pyriformis cells were first cultured in a proper medium in which R. E. tracer Ce-141, or Yb-169, or Tin-170 was spiked. It was assayed that 65% of the radioactivity localized in the cell pellicle. By a biochemical centrifuging procedure, pellicle was isolated from cilia, mitochondria etc……Membrane protein was extracted with a detergent, 2% sodium deoxycholate The compatible overlap of a protein elution peak and a radioactivity elution peak of the Sephadex G-100 gel chromatogram demonstrated the existanee of a R E. binding protein. By means of SDS-polyacrylamidegel electrophoresis a molecular weight of 54000 daltons of the protein was determined when compared with standard proteins.