Controlling and exploiting intrinsic unpaired electrons in metalloproteins
File(s)
Accepted version
Author(s)
Richardson, Katherine
Seif-Eddine, Maryam
Sills, Adam
Roessler, Maxie
Type
Journal Article
Abstract
Electron paramagnetic resonance spectroscopy encompasses a versatile set of techniques that allow detailed insight into intrinsically occurring paramagnetic centers in metalloproteins and enzymes that undergo oxidation-reduction reactions. In this chapter, we discuss the process from isolating the protein to acquiring and analyzing pulse EPR spectra, adopting a practical perspective. We start with considerations when preparing the protein sample, explain techniques and procedures available for determining the reduction potential of the redox-active center of interest and provide details on methodologies to trap a given paramagnetic state for detailed pulse EPR studies, with an emphasis on biochemical and spectroscopic tools available when multiple EPR-active species are present. We elaborate on some of the most commonly used pulse EPR techniques and the choices the user has to make, considering advantages and disadvantages and how to avoid pitfalls. Examples are provided throughout.
Date Issued
2022-04-21
Date Acceptance
2022-04-09
Citation
Methods in Enzymology, 2022, 666, pp.233-296
ISSN
0076-6879
Publisher
Elsevier
Start Page
233
End Page
296
Journal / Book Title
Methods in Enzymology
Volume
666
Copyright Statement
© 2022 Elsevier Inc. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence http://creativecommons.org/licenses/by-nc-nd/4.0/
Sponsor
The Leverhulme Trust
Engineering & Physical Science Research Council (EPSRC)
Grant Number
RPG-2018-183
EP/T031425/1
Subjects
Biochemistry & Molecular Biology
0601 Biochemistry and Cell Biology
Publication Status
Published
Date Publish Online
2022-04-21