Abstract:
:The Escherichia coli fumarate and nitrate reductase (FNR) regulator protein is an important transcriptional regulator that controls the expression of a large regulon of more than 100 genes in response to changes in oxygen availability. FNR is active when it acquires a [4Fe-4S](2+) cluster under anaerobic conditions. The presence of the [4Fe-4S](2+) cluster promotes protein dimerization and site-specific DNA binding, facilitating activation or repression of target promoters. Oxygen is sensed by the controlled disassembly of the [4Fe-4S](2+) cluster, ultimately resulting in inactive, monomeric, apo-FNR. The FNR [4Fe-4S](2+) cluster is also sensitive to nitric oxide, such that under anaerobic conditions the protein is inactivated by nitrosylation of the iron-sulfur cluster, yielding a mixture of monomeric and dimeric dinitrosyl-iron cysteine species. This chapter describes some of the methods used to produce active [4Fe-4S] FNR protein and investigates the reaction of the [4Fe-4S](2+) cluster with nitric oxide and oxygen in vitro.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Crack JC,Le Brun NE,Thomson AJ,Green J,Jervis AJdoi
10.1016/S0076-6879(07)37011-0subject
Has Abstractpub_date
2008-01-01 00:00:00pages
191-209eissn
0076-6879issn
1557-7988pii
S0076-6879(07)37011-0journal_volume
437pub_type
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