霍乱弧菌Na⁺转运NADH：泛醌氧化还原酶的NqrA和NqrC亚基的结晶及初步分析
Crystallization and preliminary analysis of the NqrA and NqrC subunits of the Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae.
作者信息
Vohl Georg, Nedielkov Ruslan, Claussen Björn, Casutt Marco S, Vorburger Thomas, Diederichs Kay, Möller Heiko M, Steuber Julia, Fritz Günter
机构信息
Institute for Neuropathology, University of Freiburg, Breisacher Strasse 64, 79106 Freiburg, Germany.
Department of Chemistry and Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, Konstanz, 78457, Germany.
出版信息
Acta Crystallogr F Struct Biol Commun. 2014 Jul;70(Pt 7):987-92. doi: 10.1107/S2053230X14009881. Epub 2014 Jun 19.
Abstract
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 Å 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 Å resolution.
摘要
霍乱弧菌的钠转运烟酰胺腺嘌呤二核苷酸（NADH）：泛醌氧化还原酶（Na⁺-NQR）是一种由六个不同亚基NqrA-NqrF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进行了异源表达并结晶。分别通过单波长反常散射（SAD）相位法和分子置换法，在空间群C222₁和P2₁中解析了NqrA1-377的结构，分辨率分别为1.9 Å和2.1 Å。去除跨膜螺旋的NqrC与ApbE共表达，以插入与苏氨酸225共价连接的黄素单核苷酸基团。以多形拟杆菌的脱辅基NqrC为搜索模型，通过分子置换法在1.8 Å分辨率下确定了该结构。
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