Pellegrini, E., Juyoux, P., von Velsen, J. et al. (6 more authors) (2024) Metal fluorides—multi-functional tools for the study of phosphoryl transfer enzymes, a practical guide. Structure, 32 (10). 1834-1846.e3. ISSN 0969-2126
Abstract
Enzymes facilitating the transfer of phosphate groups constitute the most extensive protein families across all kingdoms of life. They make up approximately 10% of the proteins found in the human genome. Understanding the mechanisms by which enzymes catalyze these reactions is essential in characterizing the processes they regulate. Metal fluorides can be used as multifunctional tools to study these enzymes. These ionic species bear the same charge as phosphate and the transferring phosphoryl group and, in addition, allow the enzyme to be trapped in catalytically important states with spectroscopically sensitive atoms interacting directly with active site residues. The ionic nature of these phosphate surrogates also allows their removal and replacement with other analogs. Here, we describe the best practices to obtain these complexes, their use in NMR, X-ray crystallography, cryo-EM, and SAXS and describe a new metal fluoride, scandium tetrafluoride, which has significant anomalous signal using soft X-rays.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2024 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | (19)F-NMR; SAXS; aluminum fluoride; cryo-EM; macromolecular crystallography; magnesium fluoride; metallofluoride; phosphoryl transfer; scandium fluoride; transition state analog |
Dates: |
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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/M021637/1 Biotechnology and Biological Sciences Research Council BB/S007695/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 21 Aug 2024 14:51 |
Last Modified: | 23 Nov 2024 05:20 |
Status: | Published |
Publisher: | Elsevier BV |
Refereed: | Yes |
Identification Number: | 10.1016/j.str.2024.07.007 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:216336 |