Brugna M, Giudici-Orticoni M T, Spinelli S, Brown K, Tegoni M, Bruschi M
Laboratoire de Bioénergétique et Ingéniérie des Protéines, C.N.R.S., Institut de Biologie Structurale et Microbiologie, Marseille, France.
Proteins. 1998 Dec 1;33(4):590-600.
Hydrogenases from Desulfovibrio are found to catalyze hydrogen uptake with low potential multiheme cytochromes, such as cytochrome c3, acting as acceptors. The production of Fe-only hydrogenase from Desulfovibrio vulgaris Hildenborough was improved with respect to the growth phase and media to determine the best large-scale bacteria growth conditions. The interaction and electron transfer from Fe-only hydrogenase to multiheme cytochrome has been studied in detail by both BLAcore and steady-state measurements. The electron transfer between [Fe] hydrogenase and cytochrome c3 appears to be a cooperative phenomenon (h = 1.37). This behavior could be related to the conductivity properties of multihemic cytochromes. An apparent dissociation constant was determined (2 x 10(-7) M). The importance of the cooperativity for contrasting models proposed to describe the functional role of the hydrogenase/cytochrome c3 complex is discussed. Presently, the only determined structure is from [NiFe] hydrogenase and there are no obvious similarities between [NiFe] and [Fe] hydrogenase. Furthermore, no crystallographic data are available concerning [Fe] hydrogenase. The first results on crystallization and X-ray crystallography are reported.
已发现来自脱硫弧菌属的氢化酶可催化以低电位多血红素细胞色素（如细胞色素c3）作为受体的吸氢反应。关于生长阶段和培养基对普通脱硫弧菌希登伯勒株仅含铁氢化酶产量进行了优化，以确定最佳的大规模细菌生长条件。通过BLAcore和稳态测量详细研究了仅含铁氢化酶与多血红素细胞色素之间的相互作用和电子转移。[Fe]氢化酶与细胞色素c3之间的电子转移似乎是一种协同现象（h = 1.37）。这种行为可能与多血红素细胞色素的导电特性有关。确定了一个表观解离常数（2×10⁻⁷ M）。讨论了协同性对于描述氢化酶/细胞色素c3复合物功能作用的不同模型的重要性。目前，唯一确定的结构来自[NiFe]氢化酶，并且[NiFe]氢化酶与[Fe]氢化酶之间没有明显的相似性。此外，尚无关于[Fe]氢化酶的晶体学数据。报道了关于结晶和X射线晶体学的初步结果。
