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Rev-Erbβ is a nuclear receptor that couples circadian rhythm, metabolism, and inflammation.1-7Heme binding to the protein modulates its function as a repressor, its stability, its ability to bind other proteins, and its activity in gas sensing.8-11Rev-Erbβ binds Fe3+-heme tighter than Fe2+-heme, suggesting its activities may be regulated by the heme redox state.9Yet, this critical role of heme redox chemistry in defining the protein’s resting state and function is unknown. We demonstrate by electrochemical and whole-cell electron paramagnetic resonance experiments that Rev-Erbβ exists in the Fe3+form within the cell essentially allowing the protein to be heme-replete even at low concentrations of labile heme in the nucleus. However, being in the Fe3+redox state contradicts Rev-Erb’s known function as a gas sensor, which dogma asserts must be a Fe2+protein This paper explains why the resting Fe3+-state is congruent both with heme-binding and cellular gas sensing. We show that the binding of CO/NO elicits a striking increase in the redox potential of the Fe3+/Fe2+couple, characteristic of an EC mechanism in which the unfavorableElectrochemical reduction of heme is coupled to the highly favorableChemical reaction of gas binding, making the reduction spontaneous. Thus, Fe3+-Rev-Erbβ remains heme-loaded, crucial for its repressor activity, and only undergoes reduction when diatomic gases are present. This work has broad implications for hemoproteins where ligand-triggered redox changes cause conformational changes influencing protein’s function or inter-protein interactions, like NCoR1 for Rev-Erbβ. This study opens up the possibility of CO/NO-mediated regulation of the circadian rhythm through redox changes in Rev-Erbβ. |
