Probing the reactivity of [4Fe-4S] fumarate and nitrate reduction (FNR) regulator with O2 and NO: Increased O2 resistance and relative specificity for NO of the [4Fe-4S] L28H FNR cluster

Jason C. Crack, Patricia Amara, Eve de Rosny, Claudine Darnault, Melanie R. Stapleton, Jeffrey Green, Anne Volbeda, Juan C. Fontecilla-Camps, Nick E. Le Brun

Research output: Contribution to journalArticlepeer-review

1 Downloads (Pure)


The Escherichia coli fumarate and nitrate reduction (FNR) regulator acts as the cell’s master switch for the transition between anaerobic and aerobic respiration, controlling the expression of >300 genes in response to O2 availability. Oxygen is perceived through a reaction with FNR’s [4Fe-4S] cluster cofactor. In addition to its primary O2 signal, the FNR [4Fe-4S] cluster also reacts with nitric oxide (NO). In response to physiological concentrations of NO, FNR de-represses the transcription of hmp, which encodes a principal NO-detoxifying enzyme, and fails to activate the expression of the nitrate reductase (nar) operon, a significant source of endogenous cellular NO. Here, we show that the L28H variant of FNR, which is much less reactive towards O2 than wild-type FNR, remains highly reactive towards NO. A high resolution structure and molecular dynamics (MD) simulations of the closely related L28H-FNR from Aliivibrio fischeri revealed decreased conformational flexibility of the Cys20-Cys29 cluster-binding loop that is suggested to inhibit outer-sphere O2 reactivity, but only partially impair inner-sphere NO reactivity. Our data provide new insights into the mechanistic basis for how iron–sulfur cluster regulators can distinguish between O2 and NO.
Original languageEnglish
Article number450
Number of pages18
Issue number12
Early online date21 Nov 2023
Publication statusPublished - Dec 2023


  • FNR
  • gene regulation
  • iron–sulfur
  • molecular dynamics
  • nitric oxide sensing

Cite this