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.
Publication types
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Research Support, Non-U.S. Gov't
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Review
MeSH terms
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Crystallography, X-Ray
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Cytochrome c Group / chemistry
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Cytochrome c Group / isolation & purification
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Cytochrome c Group / metabolism
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Cytochrome c Group / physiology*
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Cytochromes a1 / chemistry
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Cytochromes a1 / isolation & purification
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Cytochromes a1 / metabolism
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Cytochromes a1 / physiology
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Cytochromes c1 / chemistry
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Cytochromes c1 / isolation & purification
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Cytochromes c1 / metabolism
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Cytochromes c1 / physiology
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Escherichia coli / enzymology*
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Escherichia coli / growth & development
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Models, Molecular
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Nitrate Reductases / chemistry
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Nitrate Reductases / isolation & purification
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Nitrate Reductases / metabolism
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Nitrate Reductases / physiology
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Nitric Oxide / metabolism
Substances
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Cytochrome c Group
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Nitric Oxide
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Cytochromes a1
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Cytochromes c1
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cytochrome C-552
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Nitrate Reductases
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nitrate reductase (cytochrome)