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牛线粒体起始因子和延伸因子。

Bovine mitochondrial initiation and elongation factors.

作者信息

Schwartzbach C J, Farwell M, Liao H X, Spremulli L L

机构信息

Burroughs Wellcome Company. Research Triangle Park, North Carolina 27709, USA.

出版信息

Methods Enzymol. 1996;264:248-61. doi: 10.1016/s0076-6879(96)64025-7.

Abstract

The procedures summarized above provide nearly homogeneous preparations of IF-2mt, EF-Tu. Tsmt, and EF-Gmt. The scheme developed for IF-2mr leads to a 24,000-fold purification of this factor with a 26% recovery of activity. Analysis by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography indicates that this factor functions as a monomer with a molecular weight of about 85,000. The scheme developed EF-Tu.Tsmt provides a 10,000-fold purification with an overall yield of about 10%. The EF-Tumt component in this complex has a molecular weight of about 46,000, whereas EF-Tsmt has a molecular weight of about 32,000 on SDS-polyacrylamide gel electrophoresis. The EF-Tu. Tsmt complex is tightly associated and appears to have a native molecular weight of about 70,000. The five-step purification procedure outlined above for EF-Gmt results in a 14,000-fold purification of EF-Gmt with a 2-5% recovery of activity. Analysis by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography indicates that EF-Gmt functions as a monomeric protein with an apparent molecular weight of about 80,000.

摘要

上述步骤可得到近乎均一的IF-2mt、EF-Tu、Tsmt和EF-Gmt制剂。为IF-2mr制定的方案可使该因子纯化24000倍,活性回收率为26%。通过SDS-聚丙烯酰胺凝胶电泳和凝胶过滤色谱分析表明,该因子以单体形式发挥作用,分子量约为85000。为EF-Tu.Tsmt制定的方案可使其纯化10000倍,总产率约为10%。在SDS-聚丙烯酰胺凝胶电泳上,该复合物中的EF-Tumt组分分子量约为46000,而EF-Tsmt分子量约为32000。EF-Tu.Tsmt复合物紧密结合,其天然分子量似乎约为70000。上述针对EF-Gmt的五步纯化程序可使EF-Gmt纯化14000倍,活性回收率为2%-5%。通过SDS-聚丙烯酰胺凝胶电泳和凝胶过滤色谱分析表明,EF-Gmt作为一种单体蛋白发挥作用,表观分子量约为80000。

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