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Generating single metalloprotein crystals in well-defined redox states: electrochemical control combined with infrared imaging of a NiFe hydrogenase crystal.
journal contribution
posted on 2019-06-18, 08:08 authored by PA Ash, SB Carr, HA Reeve, A Skorupskaitė, JS Rowbotham, R Shutt, MD Frogley, RM Evans, G Cinque, FA Armstrong, KA VincentWe describe an approach to generating and verifying well-defined redox states in metalloprotein single crystals by combining electrochemical control with synchrotron infrared microspectroscopic imaging. For NiFe hydrogenase 1 from Escherichia coli we demonstrate fully reversible and uniform electrochemical reduction from the oxidised inactive to the fully reduced state, and temporally resolve steps during this reduction.
Funding
The work of FAA, SBC, RME and KAV is supported by BBSRC grants BB/L009722/1 and BB/N006321/1. Work of KAV, PAA, HAR and JSR is additionally supported by EPSRC grant EP/N013514/1. FAA is a Royal Society Wolfson Research Merit Award holder. AS is grateful for a scholarship from the Oxford Interdisciplinary Bioscience Doctoral Training Partnership, BB/M011224/1. We thank Diamond Light Source for access to the MIRIAM beamline B22 (SM13879) that contributed to the results presented here. We also thank Elena Nomerotskaia for assistance with isolation of Hyd1.
History
Citation
Chemical Communications, 2017, 53 (43), pp. 5858-5861Author affiliation
/Organisation/COLLEGE OF SCIENCE AND ENGINEERING/Department of ChemistryVersion
- VoR (Version of Record)
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Chemical CommunicationsPublisher
Royal Society of Chemistryeissn
1364-548XAcceptance date
2017-05-09Copyright date
2017Available date
2019-06-18Publisher DOI
Publisher version
https://pubs.rsc.org/en/content/articlelanding/2017/CC/C7CC02591B#!divAbstractNotes
Electronic supplementary information (ESI) available: Crystal preparation; cyclic voltammogram in the experimental cell; position-dependent difference spectra; demonstration of control over all active sites; visible images of a crystal recorded periodically during measurements; time-dependent Nia-R formation. See DOI: 10.1039/c7cc02591bLanguage
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