The extraction of proteins from eukaryotic ribosomes and ribosomal subunits
The extraction of proteins from eukaryotic ribosomes and ribosomal subunits
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从真核核糖体和核糖体亚基中提取蛋白质
DOI：
10.1007/bf00264778
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
I. Wool
中科院分区：
文献类型：
--
作者：
C. C. Sherton;I. Wool
从大鼠肝脏核糖体和核糖体亚基中提取蛋白质：用67%乙酸（在3.3 mM、33 mM或67 mM Mg++存在下）;用2 M LiCl的4 M尿素溶液;用0.25 N HCl;用1% SDS;以及在RNA酶消化后。最有效的提取和最好的回收率是在33 mM或67 mM Mg++存在下用乙酸，或用LiCl-尿素。用乙酸、LiCl-尿素或HCl提取的蛋白质很少或没有RNA污染。核糖体蛋白质进行了分析，通过二维聚丙烯酰胺凝胶电泳：用乙酸提取的蛋白质是最可溶的样品凝胶溶液中，其电泳图谱显示的斑点的最大数量和最小数量的衍生物或改变的蛋白质。用SDS或RNase提取的蛋白质制剂在样品凝胶溶液中相对不溶，用HCl提取的蛋白质显示出大量的衍生物。综合考虑，从真核核糖体中提取蛋白质的最令人满意的方法是在33 mM MgCl 2存在下用67%乙酸。
Proteins were extracted from rat liver ribosomes and ribosomal subunits: with 67% acetic acid (in the presence of 3.3 mM, 33 mM, or 67 mM Mg++); with 2 M LiCl in 4 M urea; with 0.25 N HCl; with 1% SDS; and after RNase digestion. The most efficient extraction and the best recovery were either with acetic acid in the presence of 33 mM or 67 mM Mg++, or with LiCl-urea. Protein extracted with acetic acid, LiCl-urea, or with HCl had little or no contamination with RNA. The ribosomal proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis: the proteins extracted with acetic acid were the most soluble in the sample gel solution; their electrophoretograms displayed the maximum number of spots and the smallest number of derivatives or altered proteins. Preparations of protein extracted with SDS or RNase were relatively insoluble in the sample gel solution, and proteins extracted with HCl showed a large number of derivatives. All things considered, the most satisfactory method for the extraction of protein from eukaryotic ribosomes is with 67% acetic acid in the presence of 33 mM MgCl2.