PMID: 2118386Aug 17, 1990Paper

Characterization of FMN-dependent NADH-quinone reductase induced by menadione in Escherichia coli

Biochimica Et Biophysica Acta
M HayashiT Unemoto

Abstract

It was found that when Escherichia coli is grown in the presence of 0.2-0.3 mM menadione (2-methyl-1,4-naphthoquinone), an FMN-dependent NADH-quinone reductase increases more than 20-fold in the cytoplasmic fraction. The menadione-induced quinone reductase was isolated from the cytoplasmic fraction of induced cells. The purified enzyme had an Mr of 24 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme required flavin as a cofactor and a half-maximum activity was obtained with 0.54 microM FMN or 16.5 microM FAD. The enzyme had a broad pH optimum at pH 7.0-8.0 and reacted with NADH, but not with NADPH. The reaction followed a ping-pong mechanism and the intrinsic Km values for NADH and menadione were estimated to be 132 microM and 2.0 microM, respectively. Dicoumarol was a simple competitive inhibitor with respect to NADH with a Ki value of 0.22 microM. The electron acceptor specificity of this enzyme was very similar to that of NAD(P)H: (quinone acceptor) oxidoreductase (EC 1.6.99.2, DT-diaphorase) from rat liver. Since menadione is reduced by the two-electron reduction pathway to menadiol, the induction of this enzyme is likely to be an adaptive response of E. coli to partially alleviate the toxicity of menadione.

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Citations

Aug 2, 2007·Journal of Structural and Functional Genomics·Allan MatteMiroslaw Cygler
Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Melanie A AdamsZongchao Jia
May 5, 2006·Journal of Bacteriology·Eric V Patridge, James G Ferry
Dec 31, 2005·Journal of Bacteriology·Jenn-Wei ChenWan-Jr Syu
Jul 10, 2010·Proceedings of the National Academy of Sciences of the United States of America·Oscar JuárezBlanca Barquera
Jul 1, 2004·Critical Reviews in Biotechnology·Zhonghu BaiBrian McNeil
Apr 5, 2014·Acta Crystallographica. Section D, Biological Crystallography·Zixi WangXiao-Dong Su
Aug 8, 2001·Archives of Biochemistry and Biophysics·M A JorgensenG L Mendz
Feb 15, 1996·Biochimica Et Biophysica Acta·M HayashiT Unemoto
Feb 8, 1993·Biochimica Et Biophysica Acta·T Yagi
Jan 14, 2009·Archives of Biochemistry and Biophysics·Vojtech SedlácekIgor Kucera

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