α-Fluorophosphonates reveal how a phosphomutase conserves transition state conformation over hexose recognition in its two-step reaction

Jin, Yi ORCID: https://orcid.org/0000-0002-6927-4371, Bhattasali, Debabrata, Pellegrini, Erika, Forget, Stephanie M., Baxter, Nicola J., Cliff, Matthew J., Bowler, Matthew W., Jakeman, David L., Blackburn, G. Michael and Waltho, Jonathan P. 2014. α-Fluorophosphonates reveal how a phosphomutase conserves transition state conformation over hexose recognition in its two-step reaction. Proceedings of the National Academy of Sciences 111 (34) , pp. 12384-12389. 10.1073/pnas.1402850111

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Abstract

β-Phosphoglucomutase (βPGM) catalyzes isomerization of β-d-glucose 1-phosphate (βG1P) into d-glucose 6-phosphate (G6P) via sequential phosphoryl transfer steps using a β-d-glucose 1,6-bisphosphate (βG16BP) intermediate. Synthetic fluoromethylenephosphonate and methylenephosphonate analogs of βG1P deliver novel step 1 transition state analog (TSA) complexes for βPGM, incorporating trifluoromagnesate and tetrafluoroaluminate surrogates of the phosphoryl group. Within an invariant protein conformation, the β-d-glucopyranose ring in the βG1P TSA complexes (step 1) is flipped over and shifted relative to the G6P TSA complexes (step 2). Its equatorial hydroxyl groups are hydrogen-bonded directly to the enzyme rather than indirectly via water molecules as in step 2. The (C)O–P bond orientation for binding the phosphate in the inert phosphate site differs by ∼30° between steps 1 and 2. By contrast, the orientations for the axial O–Mg–O alignment for the TSA of the phosphoryl group in the catalytic site differ by only ∼5°, and the atoms representing the five phosphorus-bonded oxygens in the two transition states (TSs) are virtually superimposable. The conformation of βG16BP in step 1 does not fit into the same invariant active site for step 2 by simple positional interchange of the phosphates: the TS alignment is achieved by conformational change of the hexose rather than the protein.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Chemistry
Subjects:	Q Science > QD Chemistry
Uncontrolled Keywords:	phosphonate analogs; phosphoryl transfer mechanism; 19F NMR; X-ray crystallography; water-mediated substrate recognition
Publisher:	National Academy of Sciences
ISSN:	0027-8424
Date of Acceptance:	15 July 2014
Last Modified:	06 Jan 2024 02:22
URI:	https://orca.cardiff.ac.uk/id/eprint/100301

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