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Generating single metalloprotein crystals in well-defined redox states: electrochemical control combined with infrared imaging of a NiFe hydrogenase crystal.

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posted on 18.06.2019, 08:08 by PA Ash, SB Carr, HA Reeve, A Skorupskaitė, JS Rowbotham, R Shutt, MD Frogley, RM Evans, G Cinque, FA Armstrong, KA Vincent
We 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. 5858-5861

Author affiliation

/Organisation/COLLEGE OF SCIENCE AND ENGINEERING/Department of Chemistry

Version

VoR (Version of Record)

Published in

Chemical Communications

Publisher

Royal Society of Chemistry

eissn

1364-548X

Acceptance date

09/05/2017

Copyright date

2017

Available date

18/06/2019

Publisher version

https://pubs.rsc.org/en/content/articlelanding/2017/CC/C7CC02591B#!divAbstract

Notes

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/c7cc02591b

Language

en