Active Intermediates in Copper Nitrite Reductase Reactions Probed by a Cryotrapping‐Electron Paramagnetic Resonance Approach

Tobias M. Hedison, Muralidharan Shanmugam, Derren J. Heyes, Ruth Edge, Nigel Scrutton

Research output: Contribution to journalArticlepeer-review

Abstract

Redox active metalloenzymes catalyse a range of biochemical processes essential for life. However, due to their complex reaction mechanisms, and often, their poor optical signals, detailed mechanistic understandings of them are limited. Here, we develop a cryoreduction approach coupled to electron paramagnetic resonance measurements to study electron transfer between the copper centers in the copper nitrite reductase (CuNiR) family of enzymes. Unlike alternative methods used to study electron transfer reactions, the cryoreduction approach presented here allows observation of the redox state of both metal centers, a direct read‐out of electron transfer, determines the presence of the substrate/product in the active site and shows the importance of protein motion in inter‐copper electron transfer catalyzed by CuNiRs. Cryoreduction‐EPR is broadly applicable for the study of electron transfer in other redox enzymes and paves the way to explore transient states in multiple redox‐centre containing proteins (homo and hetero metal ions).
Original languageEnglish
JournalAngewandte Chemie International Edition
Early online date30 Apr 2020
DOIs
Publication statusE-pub ahead of print - 30 Apr 2020

Research Beacons, Institutes and Platforms

  • Manchester Institute of Biotechnology
  • Dalton Nuclear Institute

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