Issue 12, 2017

Observing enzyme ternary transition state analogue complexes by 19F NMR spectroscopy

Abstract

Ternary transition state analogue (TSA) complexes probing the isomerization of β-D-glucose 1-phosphate (G1P) into D-glucose 6-phosphate (G6P) catalyzed by catalytically active, fluorinated (5-fluorotryptophan), β-phosphoglucomutase (βPGM) have been observed directly by 19F NMR spectroscopy. In these complexes MgF3 and AlF4 are surrogates for the transferring phosphate. However, the relevance of these metal fluorides as TSA complexes has been queried. The 1D 19F spectrum of a ternary TSA complex presented a molar equivalence between fluorinated enzyme, metal fluoride and non-isomerizable fluoromethylenephosphonate substrate analogue. Ring flips of the 5-fluoroindole ring remote from the active site were observed by both 19F NMR and X-ray crystallography, but did not perturb function. This data unequivocally demonstrates that the concentration of the metal fluoride complexes is equivalent to the concentration of enzyme and ligand in the TSA complex in aqueous solution.

Graphical abstract: Observing enzyme ternary transition state analogue complexes by 19F NMR spectroscopy

Supplementary files

Article information

Article type
Edge Article
Submitted
28 Sep 2017
Accepted
23 Oct 2017
First published
23 Oct 2017
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2017,8, 8427-8434

Observing enzyme ternary transition state analogue complexes by 19F NMR spectroscopy

A. Ampaw, M. Carroll, J. von Velsen, D. Bhattasali, A. Cohen, Matthew W. Bowler and D. L. Jakeman, Chem. Sci., 2017, 8, 8427 DOI: 10.1039/C7SC04204C

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. You can use material from this article in other publications, without requesting further permission from the RSC, provided that the correct acknowledgement is given and it is not used for commercial purposes.

To request permission to reproduce material from this article in a commercial publication, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party commercial publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements