Vandecasteele J P, Burris R H
J Bacteriol. 1970 Mar;101(3):794-801. doi: 10.1128/jb.101.3.794-801.1970.
A new procedure for a rapid and extensive purification of the FeMo protein and the Fe protein of the nitrogenase complex from Clostridium pasteurianum is described. Specific activities of 345 and 460 nmoles of N(2) reduced per mg of protein per min for the FeMo protein and for the Fe protein, respectively, have been obtained. Preparations of the FeMo protein contained 0.96 atom of molybdenum and 15 atoms of iron per molecule, whereas those of the Fe protein contained 2.86 atoms of iron per molecule. Experiments suggest that a definite association of two Fe proteins and one FeMo protein is functional in the active enzyme complex. No individual role could be ascribed to either of the two proteins, but the fact that hydrogenase inhibits N(2) fixation but not the reductant-dependent adenosine triphosphate hydrolysis supports the idea that there are two distinct sites on nitrogenase, one concerned with N(2) activation and the other with activated electron transport.
本文描述了一种从巴氏梭菌中快速大量纯化固氮酶复合物的铁钼蛋白和铁蛋白的新方法。分别获得了铁钼蛋白和铁蛋白的比活性，即每毫克蛋白质每分钟还原345和460纳摩尔N₂。铁钼蛋白制剂每分子含有0.96个钼原子和15个铁原子，而铁蛋白制剂每分子含有2.86个铁原子。实验表明，两个铁蛋白和一个铁钼蛋白的特定结合在活性酶复合物中起作用。两种蛋白质中的任何一种都没有明确的单独作用，但氢化酶抑制N₂固定而不抑制依赖还原剂的三磷酸腺苷水解这一事实支持了固氮酶上有两个不同位点的观点，一个与N₂活化有关，另一个与活化电子传递有关。
