Reichert G, Issinger O G
Biochim Biophys Acta. 1981 Jul 27;654(2):268-78. doi: 10.1016/0005-2787(81)90181-7.
Native 40 S particles from Krebs II mouse ascites tumor cells were isolated on a large scale. A nonribosomal protein moiety of about 30 proteins could be removed from the ribosomal particles by treatment with 250 mM KCl. These proteins were analysed by two-dimensional polyacrylamide gel electrophoresis and turned out to be mostly acidic in nature. The molecular weights of about 17 proteins were determined by three-dimensional gel electrophoresis. Radioactively labelled nonribosomal protein spots were excised from two-dimensional gel electrophoresis. Radioactively labelled nonribosomal protein spots were excised from two-dimensional gels and transferred directly or after electrodialysis onto the third dimension gel. The proteins fell into a molecular weight range from about 20,000 to 300,000.
大规模分离了来自克雷布斯II型小鼠腹水肿瘤细胞的天然40S颗粒。通过用250mM KCl处理,可以从核糖体颗粒中去除约30种蛋白质的非核糖体蛋白部分。通过二维聚丙烯酰胺凝胶电泳分析这些蛋白质,结果表明它们在性质上大多是酸性的。通过三维凝胶电泳测定了约17种蛋白质的分子量。从二维凝胶电泳中切下放射性标记的非核糖体蛋白斑点。从二维凝胶中切下放射性标记的非核糖体蛋白斑点,直接或在电渗析后转移到第三维凝胶上。这些蛋白质的分子量范围约为20,000至300,000。