Yoder M, Torres C, Corelli R, Jurnak F
Anal Biochem. 1985 Oct;150(1):243-8. doi: 10.1016/0003-2697(85)90466-x.
A simple procedure for the preparation of 10-500 mg of the Escherichia coli elongation Tu-Ts complex is described. The protocol is based on the separate purification and quantitation of EF-Tu-GDP and EF-Ts, followed by mixing of equimolar amounts of each protein and removal of the displaced GDP by dialysis. Single crystals grown from the final product have been analyzed by X-ray diffraction techniques. The procedure is also applicable to the bulk preparation and crystallization of the trypsin-modified Tu-Ts complex. Quantitation of the elongation factors by three methods is presented.
本文描述了一种制备10 - 500毫克大肠杆菌延伸因子Tu - Ts复合物的简单方法。该方案基于分别纯化和定量EF - Tu - GDP和EF - Ts,然后将等摩尔量的每种蛋白质混合,并通过透析去除置换出的GDP。通过X射线衍射技术分析了从最终产物中生长出的单晶。该方法也适用于胰蛋白酶修饰的Tu - Ts复合物的大量制备和结晶。文中介绍了用三种方法对延伸因子进行定量。