Improvement in protein HSQC spectra from addition of betaine

Abstract

Addition of glycine betaine up to 1 M gave rise to increased intensity for some weak signals in the HSQC spectra of barnase and Plasmodium falciparum flap endonuclease. The signals that were enhanced were low intensity signals, often from amide groups with rapid internal motion (low order parameter). The majority of signals are however somewhat weaker because of the increased viscosity. Addition of betaine is shown to cause a small but significant overall increase in order parameter, no consistent effect on conformational change on the µs-ms timescale, and a reduction in amide exchange rates, by a factor of about 3. The results are consistent with a model in which betaine leads to a reduction in fluctuations within the bulk water, which in turn produces a reduction in internal fluctuations of the protein.

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Item Type:	Article
Authors/Creators:	O’Dea, F. Seargeant, A.J. Hurcum, J. do Aido-Machado, R. https://orcid.org/0000-0001-6331-4216 Rowe, M.L. Baxter, N.J. https://orcid.org/0009-0000-8292-7800 Waltho, J.P. Sayers, J.R. https://orcid.org/0000-0002-5082-1443 Williamson, M.P. https://orcid.org/0000-0001-5572-1903
Copyright, Publisher and Additional Information:	© The Author(s) 2025. 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:	Amide exchange; Barnase; Betaine; Dynamics; Flap endonuclease; Fluctuation; Osmolyte
Dates:	Published: 10 March 2025 Published (online): 10 March 2025 Accepted: 20 February 2025 Submitted: 9 January 2025
Institution:	The University of Sheffield
Academic Units:	The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Medicine and Population Health The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) > Department of Molecular Biology and Biotechnology (Sheffield)
Depositing User:	Symplectic Sheffield
Date Deposited:	14 Mar 2025 11:15
Last Modified:	14 Mar 2025 11:15
Status:	Published online
Publisher:	Springer Science and Business Media LLC
Refereed:	Yes
Identification Number:	10.1007/s10858-025-00463-0
Related URLs:	PubMed URL
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:224420

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Improvement in protein HSQC spectra from addition of betaine

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