Amphipathic C-terminal region of Escherichia coli NADH dehydrogenase-2 mediates membrane localization

被引：15

作者：

Villegas, Josefina M.

Volentini, Sabrina I.

Rintoul, Maria R.

Rapisarda, Viviana A. ^{[1
]}

机构：

[1] Univ Nacl Tucuman, Dept Bioquim Nutr, Inst Super Invest Biol, Consejo Nacl Invest Cient & Tecn, San Miguel De Tucuman, Argentina

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 2011年 / 505卷 / 02期

关键词：

NADH:quinone oxidoreductase; Type 2 NADH dehydrogenase; FAD; Quinones; Membrane proteins; Respiratory chain; QUINONE OXIDOREDUCTASE; UBIQUINONE OXIDOREDUCTASE; SACCHAROMYCES-CEREVISIAE; BINDING-SITE; PURIFICATION; MITOCHONDRIA; IDENTIFICATION; REDUCTASE; COMPLEX; ENZYME;

D O I：

10.1016/j.abb.2010.10.001

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Respiratory NADH dehydrogenase-2 (NDH-2) of Escherichia coli is a membrane-bound flavoprotein. Bio-informatics approaches suggested the involvement of NDH-2 C-terminal region in membrane anchorage. Here, we demonstrated that NDH-2 is a peripheral membrane protein and that its predicted C-terminal amphipathic Arg390-Ala406 helix is sufficient to bind the protein to lipid membranes. Additionally, a cytosolic NDH-2 protein (Trun-3), lacking the last 43 aminoacids, was purified and characterized. FAD cofactor was absent in purified Trun-3. Upon the addition of FAD, Trun-3 maximum velocity was similar to native NDH-2 rate with ferricyanide and MU acceptors. However, Trun-3 activity was around 5-fold lower with quinones. No significant difference in Km values was observed for both enzymes. For the first time, an active and water soluble NDH-2 was obtained, representing a major improvement for structural/functional characterizations. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：155 / 159

页数：5

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