PMID: 9428711Jan 15, 1998Paper

Kinetics of membrane-bound nitrate reductase A from Escherichia coli with analogues of physiological electron donors--different reaction sites for menadiol and duroquinol

European Journal of Biochemistry
Roger GiordaniM L Cárdenas

Abstract

We have compared the steady-state kinetics of wild-type nitrate reductase A and two mutant forms with altered beta subunits. To mimic conditions in vivo as closely as possible, we used analogues of the physiological quinols as electron donors and membranes with overexpressed nitrate reductase A in preference to a purified alpha beta gamma complex. With the wild-type enzyme both menadiol and duroquinol supply their electrons for the reduction of nitrate at rates that depend on the square of the quinol concentration, menadiol having the higher catalytic constant. The results as a whole are consistent with a substituted-enzyme mechanism for the reduction of nitrate by the quinols. Kinetic experiments suggest that duroquinol and menadiol deliver their electrons at different sites on nitrate reductase, with cross-inhibition. Menadiol inhibits the duroquinol reaction strongly, suggesting that menaquinol may be the preferred substrate in vivo. To examine whether electron transfer from menadiol and duroquinol for nitrate reduction requires the presence of all of the Fe-S centres, we have studied the steady-state kinetics of mutants with beta subunits that lack an Fe-S centre. The loss of the highest-potential Fe-S centre results in an ...Continue Reading

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Citations

Apr 27, 2011·Proceedings of the National Academy of Sciences of the United States of America·Rodrigo Arias-CartinAxel Magalon
Mar 1, 2005·Journal of Biochemical and Molecular Toxicology·Gabriela ChaufanMaría Del Carmen Ríos de Molina
Feb 5, 2013·Biochimica Et Biophysica Acta·Stéphane GrimaldiAxel Magalon
Nov 7, 2009·The Journal of Biological Chemistry·Stéphane GrimaldiBruno Guigliarelli
Jun 16, 2019·Environmental Science and Pollution Research International·Sahar A ZakiDesouky A Abd-El-Haleem
Sep 30, 2008·The Biochemical Journal·Sophie A WeissLars J C Jeuken
Apr 15, 2010·Journal of the American Chemical Society·Rodrigo Arias-CartinStéphane Grimaldi
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·M K Berlyn
Jun 25, 2014·Biochemistry·Justin G FedorJoel H Weiner

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