Principles and practice of determining metal–protein affinities

Young, Tessa R.; Xiao, Zhiguang

doi:10.1042/bcj20200838

Principles and practice of determining metal–protein affinities

Young, Tessa R.; Xiao, Zhiguang

Authors

Dr Tessa Young tessa.r.young@durham.ac.uk
Assistant Professor

Zhiguang Xiao

Abstract

Metal ions play many critical roles in biology, as structural and catalytic cofactors, and as cell regulatory and signalling elements. The metal–protein affinity, expressed conveniently by the metal dissociation constant, KD, describes the thermodynamic strength of a metal–protein interaction and is a key parameter that can be used, for example, to understand how proteins may acquire metals in a cell and to identify dynamic elements (e.g. cofactor binding, changing metal availabilities) which regulate protein metalation in vivo. Here, we outline the fundamental principles and practical considerations that are key to the reliable quantification of metal–protein affinities. We review a selection of spectroscopic probes which can be used to determine protein affinities for essential biological transition metals (including Mn(II), Fe(II), Co(II), Ni(II), Cu(I), Cu(II) and Zn(II)) and, using selected examples, demonstrate how rational probe selection combined with prudent experimental design can be applied to determine accurate KD values.

Citation

Young, T. R., & Xiao, Z. (2021). Principles and practice of determining metal–protein affinities. Biochemical Journal, 478(5), 1085-1116. https://doi.org/10.1042/bcj20200838

Journal Article Type	Article
Acceptance Date	Feb 11, 2021
Online Publication Date	Mar 10, 2021
Publication Date	2021
Deposit Date	Mar 13, 2021
Publicly Available Date	May 6, 2021
Journal	Biochemical Journal
Print ISSN	0264-6021
Electronic ISSN	1470-8728
Publisher	Portland Press
Peer Reviewed	Peer Reviewed
Volume	478
Issue	5
Pages	1085-1116
DOI	https://doi.org/10.1042/bcj20200838

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Copyright Statement
© 2021 The Author(s). This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY)