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 language | English |
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Pages (from-to) | 63-77 |
Number of pages | 15 |
Journal | Methods in Enzymology |
Volume | 437 |
DOIs | |
Publication status | Published - 2008 |