Metal fluorides-multi-functional tools for the study of phosphoryl transfer enzymes, a practical guide.

Autor:	Pellegrini E; European Molecular Biology Laboratory, 71 avenue des Martyrs, CS 90181, 38042 Grenoble, France., Juyoux P; European Molecular Biology Laboratory, 71 avenue des Martyrs, CS 90181, 38042 Grenoble, France., von Velsen J; European Molecular Biology Laboratory, 71 avenue des Martyrs, CS 90181, 38042 Grenoble, France., Baxter NJ; School of Biosciences, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK., Dannatt HRW; School of Biosciences, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK., Jin Y; School of Biosciences, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK., Cliff MJ; Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK., Waltho JP; School of Biosciences, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK; Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK. Electronic address: j.waltho@manchester.ac.uk., Bowler MW; European Molecular Biology Laboratory, 71 avenue des Martyrs, CS 90181, 38042 Grenoble, France. Electronic address: mbowler@embl.fr.
Jazyk:	angličtina
Zdroj:	Structure (London, England : 1993) [Structure] 2024 Oct 03; Vol. 32 (10), pp. 1834-1846.e3. Date of Electronic Publication: 2024 Aug 05.
DOI:	10.1016/j.str.2024.07.007
Abstrakt:	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. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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