Reversible cycling between cysteine persulfide-ligated [2Fe-2S] and cysteine-ligated [4Fe-4S] clusters in the FNR regulatory protein

Bo Zhang, Jason C. Crack, Sowmya Subramanian, Jeffrey Green, Andrew J. Thomson, Nick E. Le Brun, Michael K. Johnson

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Fumarate and nitrate reduction (FNR) regulatory proteins are O2-sensing bacterial transcription factors that control the switch between aerobic and anaerobic metabolism. Under anaerobic conditions [4Fe-4S]2+-FNR exists as a DNA-binding homodimer. In response to elevated oxygen levels, the [4Fe-4S]2+ cluster undergoes a rapid conversion to a [2Fe-2S]2+ cluster, resulting in a dimer-to-monomer transition and loss of site-specific DNA binding. In this work, resonance Raman and UV-visible absorption/CD spectroscopies and MS were used to characterize the interconversion between [4Fe-4S]2+ and [2Fe-2S]2+ clusters in Escherichia coli FNR. Selective 34S labeling of the bridging sulfides in the [4Fe-4S]2+ cluster-bound form of FNR facilitated identification of resonantly enhanced Cys32S-34S stretching modes in the resonance Raman spectrum of the O2-exposed [2Fe-2S]2+ cluster-bound form of FNR. This result indicates O2-induced oxidation and retention of bridging sulfides in the form of [2Fe-2S]2+ cluster-bound cysteine persulfides. MS also demonstrates that multiple cysteine persulfides are formed on O2 exposure of [4Fe-4S]2+-FNR. The [4Fe-4S]2+ cluster in FNR can also be regenerated from the cysteine persulfide-coordinated [2Fe-2S]2+ cluster by anaerobic incubation with DTT and Fe2+ ion in the absence of exogenous sulfide. Resonance Raman data indicate that this type of cluster conversion involving sulfide oxidation is not unique to FNR, because it also occurs in O2-exposed forms of O2-sensitive [4Fe-4S] clusters in radical S-adenosylmethionine enzymes. The results provide fresh insight into the molecular mechanism of O2 sensing by FNR and iron-sulfur cluster conversion reactions in general, and suggest unique mechanisms for the assembly or repair of biological [4Fe-4S] clusters.
Original languageEnglish
Pages (from-to)15734-15739
Number of pages6
JournalProceedings of the National Academy of Sciences
Volume109
Issue number39
DOIs
Publication statusPublished - 25 Sep 2012

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