Crystallization and preliminary structure determination of the membrane-bound complex cytochrome c nitrite reductase from Desulfovibrio vulgaris Hildenborough
Crystallization and preliminary structure determination of the membrane-bound complex cytochrome c nitrite reductase from Desulfovibrio vulgaris Hildenborough
复制标题
DOI：
10.1107/s1744309106016629
复制
发表时间：
2006-06-01
影响因子：
0.9
通讯作者：
Archer, M.
中科院分区：
文献类型：
--
作者：
Rodrigues, M. L.;Oliveira, T.;Archer, M.
从Desulfovibrio vulgaris Hildenborough中分离的细胞色素c亚硝酸盐还原酶（cytochrome c亚硝酸盐reductase， cNiR）是由NrfA和NrfH亚基组成的膜结合复合物。催化亚基NrfA是一种可溶性的五血红素细胞色素c，形成约120 kDa的生理二聚体。电子供体亚基NrfH是一种分子量约为18 kDa的膜锚定的四血红素细胞色素c，属于醌脱氢酶的NapC/NirT家族，其结构尚不清楚。以PEG 4K为沉淀剂，用十二烷基麦芽糖苷洗涤剂（DDM）溶解得到天然cNiR膜配合物晶体。在瑞士光源下以2.3埃的分辨率测量了异常衍射数据。晶体属于正交空间群P2(1)2(1)2(1)，单位胞参数a = 79.5, b = 256.7, c = 578.2埃。结合分子置换和MAD方法对其结构进行求解。数据显示，D. vulgaris cNiR每个NrfA二聚体含有一个NrfH亚基。
The cytochrome c nitrite reductase (cNiR) isolated from Desulfovibrio vulgaris Hildenborough is a membrane-bound complex formed of NrfA and NrfH subunits. The catalytic subunit NrfA is a soluble pentahaem cytochrome c that forms a physiological dimer of about 120 kDa. The electron-donor subunit NrfH is a membrane-anchored tetrahaem cytochrome c of about 18 kDa molecular weight and belongs to the NapC/NirT family of quinol dehydrogenases, for which no structures are known. Crystals of the native cNiR membrane complex, solubilized with dodecylmaltoside detergent (DDM), were obtained using PEG 4K as precipitant. Anomalous diffraction data were measured at the Swiss Light Source to 2.3 angstrom resolution. Crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 79.5, b = 256.7, c = 578.2 angstrom. Molecular-replacement and MAD methods were combined to solve the structure. The data presented reveal that D. vulgaris cNiR contains one NrfH subunit per NrfA dimer.