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

• Published in: Structure (London) Vol. 32; no. 10; pp. 1834 - 1846.e3
• Main Authors: Pellegrini, Erika, Juyoux, Pauline, von Velsen, Jill, Baxter, Nicola J., Dannatt, Hugh R.W., Jin, Yi, Cliff, Matthew J., Waltho, Jonathan P., Bowler, Matthew W.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 03.10.2024
• Subjects: 19F-NMR; aluminum fluoride; cryo-EM; macromolecular crystallography; magnesium fluoride; metallofluoride; phosphoryl transfer; SAXS; scandium fluoride; transition state analog; 19F-NMR; metallofluoride; aluminum fluoride; scandium fluoride; magnesium fluoride; macromolecular crystallography; phosphoryl transfer; transition state analog; cryo-EM; SAXS; F-NMR
• ISSN: 0969-2126; 1878-4186; 1878-4186
• DOI: 10.1016/j.str.2024.07.007

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. [Display omitted] •Phosphoryl transfer enzymes have many crucial cellular functions•Metal fluorides stabilize active conformations and are spectroscopically sensitive•Metal fluorides mimic transition and ground states•Guidelines for the formation of these complexes and their use are provided Metal fluorides mimic phosphate groups, stabilize active protein conformations, and are spectroscopically active. Pellegrini et al. present best practices for the formation and use of metal fluorides in the study of phosphoryl transfer proteins. They are useful in various techniques, including NMR, X-ray crystallography, cryo-EM, and SAXS.