The measurement of binding affinities by NMR chemical shift perturbation

Abstract

We have carried out chemical shift perturbation titrations on three contrasting proteins. The resulting chemical shifts have been analysed to determine the best way to fit the data, and it is concluded that a simultaneous fitting of all raw shift data to a single dissociation constant is both the most accurate and the most precise method. It is shown that the optimal weighting of 15N chemical shifts to 1H chemical shifts is protein dependent, but is around the consensus value of 0.14. We show that chemical shift changes of individual residues can be fit to give residue-specific affinities. Residues with affinities significantly stronger than average are found in close contact with the ligand and are suggested to form a rigid contact surface, but only when the binding involves little conformational change. This observation may be of value in analysing binding and conformational change.

Metadata

Item Type:	Article
Authors/Creators:	Hobbs, B. Drant, J. Williamson, M.P. https://orcid.org/0000-0001-5572-1903
Copyright, Publisher and Additional Information:	© The Author(s) 2022. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Keywords:	Affinity; Binding; Dissociation constant; NMR; Chemical shift; Conformational change
Dates:	Published: August 2022 Published (online): 3 August 2022 Accepted: 20 July 2022
Institution:	The University of Sheffield
Academic Units:	The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield)
Funding Information:	Funder Grant number BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL BB/R000727/1 ENGINEERING AND PHYSICAL SCIENCE RESEARCH COUNCIL EP/S01358X/1
Depositing User:	Symplectic Sheffield
Date Deposited:	22 Sep 2022 12:01
Last Modified:	10 Feb 2023 14:14
Status:	Published
Publisher:	Springer Science and Business Media LLC
Refereed:	Yes
Identification Number:	10.1007/s10858-022-00402-3
Related URLs:	Author
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:190658

Download

Published Version

Filename: s10858-022-00402-3.pdf

Licence: CC-BY 4.0

CLICK TO DOWNLOAD

CORE (COnnecting REpositories)

The measurement of binding affinities by NMR chemical shift perturbation

Abstract

Metadata

Download

Published Version

Export

Statistics