title: Escherichia coli cytochrome c nitrite reductase NrfA.
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.
authors.affiliation: Centre for Metalloprotein Spectroscopy and Biology, School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Clarke
authors.forename: Paul C
authors.initials: PC
authors.lastname: Mills
authors.forename: Susie R
authors.initials: SR
authors.lastname: Poock
authors.forename: Julea N
authors.initials: JN
authors.lastname: Butt
authors.forename: Myles R
authors.initials: MR
authors.lastname: Cheesman
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Cole
authors.forename: Jay C D
authors.initials: JC
authors.lastname: Hinton
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Hemmings
authors.forename: Gemma
authors.initials: G
authors.lastname: Kemp
authors.forename: Christopher A G
authors.initials: CA
authors.lastname: Söderberg
authors.forename: Stephen
authors.initials: S
authors.lastname: Spiro
authors.forename: Jessica
authors.initials: J
authors.lastname: Van Wonderen
authors.forename: David J
authors.initials: DJ
authors.lastname: Richardson
mesh_terms: D018360:Crystallography, X-Ray; D003574:Cytochrome c Group / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D045263:Cytochromes a1 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D003575:Cytochromes c1 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D008958:Models, Molecular; D009565:Nitrate Reductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D009569:Nitric Oxide / Q000378:metabolism
chemical_list: D003574:Cytochrome c Group; D009569:Nitric Oxide; D045263:Cytochromes a1; D003575:Cytochromes c1; C022302:cytochrome C-552; D009565:Nitrate Reductases; C054690:nitrate reductase (cytochrome)