Principles and practice of determining metal–protein affinities

Young, Tessa R.; Xiao, Zhiguang

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
• Public URL: https://durham-repository.worktribe.com/output/1251309