摘要
棕色固氮菌固氮酶ＭｏＦｅ蛋白经邻菲罗啉在厌氧和有氧条件下处理后，成为部分缺失ＦｅＭｏｃｏ和Ｐ－ｃｌｕｓｔｅｒ的失活蛋白，用由钒酸钠，高柠檬酸铁、Ｎａ２Ｓ和二硫硫苏糖醇组成的溶液保温后，可使这失活蛋白变为重组蛋白。重组蛋白的ＣＤ谱除５５０－６５０ｎｍ与ＭｏＦｅ蛋白存在差异外，也能与其吸收光谱和对质子还原活性一样得到基本恢复，但对Ｃ２Ｈ２的还原活性却未得到明显恢复。表明。
By treating the reduced MoFe protein of nitrogenase from Azotobacter vinelandii with o-phenanthroline under anaerobic or aerobic condition, inactive MoFe protein which was par-tialy deficient in both P-cluster and FeMoco could be obtained. After incubating the inactive MoFe protein with a reconstituent solution containing NaVO_3, ferric homocitrate, Na_2S and dithiothreitol, a reconstituted protein could be obtained. The proton reduction activity and absorption spectrum of the reconstituted protein could be well restored, but its C_2H_2-reduc-tion activity could not be recovered. Its CD spectrum could be recovered except for the 550nm to 650nm region which differed from that of the reduced MoFe protein. The results showed that the reconstituted protein was different from MoFe protein,but was similar to vanadium-iron protein.
基金
国家基础性研究重大关键项目和国家自然科学基金项目
关键词
固氮酶
钒铁蛋白
棕色固氮菌
Nitrogenase vanadium-iron protein
Azotobacter vinelandii
Reconstituted protein
Assembly in vitro
