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Escherichia coli cytochrome c nitrite reductase NrfA. 2008
Thomas A Clarke, and
Paul C Mills, and
Susie R Poock, and
Julea N Butt, and
Myles R Cheesman, and
Jeffrey A Cole, and
Jay C D Hinton, and
Andrew M Hemmings, and
Gemma Kemp, and
Christopher A G Söderberg, and
Stephen Spiro, and
Jessica Van Wonderen, and
David J Richardson
Centre for Metalloprotein Spectroscopy and Biology, School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.
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.
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