Escherichia coli cytochrome c nitrite reductase NrfA.
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.
Clarke
Thomas A
TA
Centre for Metalloprotein Spectroscopy and Biology, School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.
Mills
Paul C
PC
Poock
Susie R
SR
Butt
Julea N
JN
Cheesman
Myles R
MR
Cole
Jeffrey A
JA
Hinton
Jay C D
JC
Hemmings
Andrew M
AM
Kemp
Gemma
G
Söderberg
Christopher A G
CA
Spiro
Stephen
S
Van Wonderen
Jessica
J
Richardson
David J
DJ
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
D003574:Cytochrome c Group; D009569:Nitric Oxide; D045263:Cytochromes a1; D003575:Cytochromes c1; C022302:cytochrome C-552; D009565:Nitrate Reductases; C054690:nitrate reductase (cytochrome)