Crystallization and preliminary analysis of the NqrA and NqrC subunits of the Na+- translocating NADH: ubiquinone oxidoreductase from Vibrio cholerae
Crystallization and preliminary analysis of the NqrA and NqrC subunits of the Na+- translocating NADH: ubiquinone oxidoreductase from Vibrio cholerae
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DOI：
10.1107/s2053230x14009881
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发表时间：
2014-07-01
影响因子：
0.9
通讯作者：
Fritz, Guenter
中科院分区：
文献类型：
--
作者：
Vohl, Georg;Nedielkov, Ruslan;Fritz, Guenter
Na+转运的NADH：霍乱弧菌泛醌氧化还原酶（Na+-NQR）是由NqrA-NqrF六个亚基组成的膜蛋白复合物。NqrA和NqrC亚基的主要结构域在大肠杆菌中异源表达并结晶。通过SAD定相和分子置换分别在1.9和2.1埃分辨率下解析了空间群C2221和P21中的NqrA 1 -377的结构。无跨膜螺旋的NqrC与ApbE共表达以插入共价连接至Thr 225的黄素单核苷酸肽基团。使用Parabacteroides distasonis的apo-NqrC作为1.8埃分辨率的搜索模型，通过分子置换确定结构。
The Na+-translocating NADH: ubiquinone oxidoreductase (Na+-NQR) from Vibrio cholerae is a membrane protein complex consisting of six different subunits NqrA-NqrF. The major domains of the NqrA and NqrC subunits were heterologously expressed in Escherichia coli and crystallized. The structure of NqrA1-377 was solved in space groups C2221 and P21 by SAD phasing and molecular replacement at 1.9 and 2.1 angstrom resolution, respectively. NqrC devoid of the transmembrane helix was co-expressed with ApbE to insert the flavin mononucleotide group covalently attached to Thr225. The structure was determined by molecular replacement using apo-NqrC of Parabacteroides distasonis as search model at 1.8 angstrom resolution.