Maier R J, Mutaftschiev S
J Biol Chem. 1982 Feb 25;257(4):2092-6.
An in vitro reconstitution of both methylene blue and oxygen-dependent H2 uptake activity from extracts of Hup- (H2 uptake-negative) mutant strains of Rhizobium japonicum bacteroids is described. Cell-free extracts prepared from bacteroids formed from two different Hup- mutants were mixed, and active H2 oxidizing particles formed. Extracts from each mutant alone did not oxidize H2. The source of the components required for the complementation were soluble. Mixing of membrane particles from the two mutants did not result in reconstituted activity. The development of activity required an incubation period of several hours under anaerobic conditions, and maximal activity was obtained approximately 10 h after the mixing of the two extracts. Along with the development of H2 uptake activity with time, the soluble extract mixture became turbid. The turbidity could be correlated with an increase in the appearance of membrane structures, including closed vesicles. After reconstitution, 65% of the methylene blue-dependent H2 uptake activity was recovered in a particulate fraction.
本文描述了从日本根瘤菌类菌体的Hup-（氢气摄取阴性）突变菌株提取物中对亚甲基蓝和氧气依赖性氢气摄取活性进行的体外重建。将由两种不同的Hup-突变体形成的类菌体制备的无细胞提取物混合，形成了具有活性的氢气氧化颗粒。单独来自每个突变体的提取物都不能氧化氢气。互补所需成分的来源是可溶的。将两个突变体的膜颗粒混合不会产生重建活性。活性的发展需要在厌氧条件下孵育数小时，并且在两种提取物混合后约10小时获得最大活性。随着氢气摄取活性随时间的发展，可溶提取物混合物变得浑浊。浑浊与包括封闭囊泡在内的膜结构外观的增加相关。重建后，65%的亚甲基蓝依赖性氢气摄取活性在颗粒部分中得以恢复。
