Escherichia coli cytochrome c nitrite reductase NrfA

Thomas A. Clarke, Paul C. Mills, Susie R. Poock, Julea N. Butt, Myles R. Cheesman, Jeffrey A. Cole, Jay C. D. Hinton, Andrew M. Hemmings, Gemma Kemp, Christopher A. G. Söderberg, Stephen Spiro, Jessica Van Wonderen, David J. Richardson

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Abstract

The periplasmic cytochrome c nitrite reductase (Nrf) system of Escherichia coli utilizes nitrite as a respiratory electron acceptor by reducing it to ammonium. Nitric oxide (NO) is a proposed intermediate in this six-electron reduction and NrfA can use exogenous NO as a substrate. This chapter describes the method used to assay Nrf-catalyzed NO reduction in whole cells of E. coli and the procedures for preparing highly purified NrfA suitable for use in kinetic, spectroscopic, voltammetric, and crystallization studies.
Original languageEnglish
Pages (from-to)63-77
Number of pages15
JournalMethods in Enzymology
Volume437
DOIs
Publication statusPublished - 2008

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