Zhu P P, Lecchi P, Pannell L, Jaffe H, Peterkofsky A
National Institutes of Health, Bethesda, Maryland 20852, USA.
Protein Expr Purif. 1995 Apr;6(2):189-95. doi: 10.1006/prep.1995.1024.
The gene (ptsH) for the phosphocarrier protein, HPr, of the phosphoenolpyruvate:sugar phosphotransferase system from Mycoplasma capricolum was previously cloned and sequenced. We present here the results of experiments in which the ptsH gene was cloned into a vector for high level expression in Escherichia coli of the phosphocarrier protein. Conditions were developed for overproduction and purification of HPr by a two-column procedure. The purified protein, analyzed by Edman degradation and mass spectrometry, was found to have been processed by removal of the N-terminal methionine residue. Examination of the purified protein by gel electrophoresis under isoelectric focusing conditions revealed that it has an unusually high isoelectric point.
糖磷酸转移酶系统的磷酸载体蛋白HPr的基因（ptsH）。我们在此展示了将ptsH基因克隆到一个载体中，以便在大肠杆菌中高水平表达磷酸载体蛋白的实验结果。通过双柱法开发了过量生产和纯化HPr的条件。经埃德曼降解和质谱分析的纯化蛋白被发现已通过去除N端甲硫氨酸残基进行了加工。在等电聚焦条件下通过凝胶电泳对纯化蛋白进行检测，结果显示其具有异常高的等电点。
