Volume 243, 2023

Electrochemical experiments define potentials associated with binding of substrates and inhibitors to nitrogenase MoFe protein

Abstract

Nitrogenases catalyse the 6-electron reduction of dinitrogen to ammonia, passing through a series of redox and protonation levels during catalytic substrate reduction. The molybdenum–iron nitrogenase is the most well-studied, but redox potentials associated with proton-coupled transformations between the redox levels of the catalytic MoFe protein have proved difficult to pin down, in part due to a complex electron-transfer pathway from the partner Fe protein, linked to ATP-hydrolysis. Here, we apply electrochemical control to the MoFe protein of Azotobacter vinelandii nitrogenase, using europium(III/II)-ligand couples as low potential redox mediators. We combine insight from the electrochemical current response with data from gas chromatography and in situ infrared spectroscopy, in order to define potentials for the binding of a series of inhibitors (carbon monoxide, methyl isocyanide) to the metallo-catalytic site of the MoFe protein, and the onset of catalytic transformation of alternative substrates (protons and acetylene) by the enzyme. Thus, we associate potentials with the redox levels for inhibition and catalysis by nitrogenase, with relevance to the elusive mechanism of biological nitrogen fixation.

Graphical abstract: Electrochemical experiments define potentials associated with binding of substrates and inhibitors to nitrogenase MoFe protein

Associated articles

Supplementary files

Article information

Article type
Paper
Submitted
27 nov 2022
Accepted
06 feb 2023
First published
06 feb 2023
This article is Open Access
Creative Commons BY license

Faraday Discuss., 2023,243, 270-286

Electrochemical experiments define potentials associated with binding of substrates and inhibitors to nitrogenase MoFe protein

T. Chen, P. A. Ash, L. C. Seefeldt and K. A. Vincent, Faraday Discuss., 2023, 243, 270 DOI: 10.1039/D2FD00170E

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