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


Journal

Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543

Informations de publication

Date de publication:
10 08 2020
Historique:
received: 07 04 2020
pubmed: 1 5 2020
medline: 17 3 2021
entrez: 1 5 2020
Statut: ppublish

Résumé

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-center containing proteins (homo and hetero metal ions).

Identifiants

pubmed: 32352195
doi: 10.1002/anie.202005052
pmc: PMC7497095
doi:

Substances chimiques

Nitrite Reductases EC 1.7.-
nitrite reductase, copper-containing EC 1.7.2.1

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

13936-13940

Subventions

Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/N013980/1
Pays : United Kingdom

Informations de copyright

© 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

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Auteurs

Tobias M Hedison (TM)

Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.
BBSRC and EPSRC funded Future Biomanfacturing Research Hub, Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.

Muralidharan Shanmugam (M)

Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.

Derren J Heyes (DJ)

Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.

Ruth Edge (R)

Dalton Cumbrian Facility, The University of Manchester, Cumbria, UK.

Nigel S Scrutton (NS)

Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.
BBSRC and EPSRC funded Future Biomanfacturing Research Hub, Manchester Institute of Biotechnology and School of Chemistry, University of Manchester, Princess Street, Manchester, M1 7DN, UK.

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Classifications MeSH