Crystallization and preliminary analysis of the NqrA and NqrC subunits of the Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae

霍乱弧菌 Na+ 转运 NADH:泛醌氧化还原酶的 NqrA 和 NqrC 亚基的结晶和初步分析

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作者:Georg Vohl, Ruslan Nedielkov, Björn Claussen, Marco S Casutt, Thomas Vorburger, Kay Diederichs, Heiko M Möller, Julia Steuber, Günter Fritz

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 C222&sub1; and P2&sub1; 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.

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