Trapping and structural characterisation of a covalent intermediate in vitamin B6 biosynthesis catalysed by the Pdx1 PLP synthase

Matthew J. Rodrigues; Nitai Giri; Antoine Royant; Yang Zhang; Rachel Bolton; Gwyndaf Evans; Steve E. Ealick; Tadhg Begley; Ivo Tews

doi:10.1039/D1CB00160D

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Trapping and structural characterisation of a covalent intermediate in vitamin B₆ biosynthesis catalysed by the Pdx1 PLP synthase†

Matthew J. Rodrigues,

^ab Nitai Giri,

^c Antoine Royant,^de Yang Zhang,^f Rachel Bolton,^ab Gwyndaf Evans,^b Steve E. Ealick,

^f Tadhg Begley^c and Ivo Tews

*^a

Author affiliations

* Corresponding authors

^a Biological Sciences, Institute for Life Sciences, University of Southampton, Southampton, UK
E-mail: ivo.tews@soton.ac.uk

^b Diamond Light Source, Harwell Science and Innovation Campus, Didcot, UK

^c Department of Chemistry, Texas A&M University, College Station, TX 77843, USA

^d Université Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale (IBS), CS 10090, Grenoble Cedex 9 38044, France

^e European Synchrotron Radiation Facility, CS 40220, Grenoble Cedex 9 38043, France

^f Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA

Abstract

The Pdx1 enzyme catalyses condensation of two carbohydrates and ammonia to form pyridoxal 5-phosphate (PLP) via an imine relay mechanism of carbonyl intermediates. The I₃₃₃ intermediate characterised here using structural, UV-vis absorption spectroscopy and mass spectrometry analyses rationalises stereoselective deprotonation and subsequent substrate assisted phosphate elimination, central to PLP biosynthesis.

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Article information

DOI: https://doi.org/10.1039/D1CB00160D
Article type: Communication
Submitted: 06 Aug 2021
Accepted: 25 Oct 2021
First published: 25 Oct 2021
This article is Open Access

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RSC Chem. Biol., 2022,3, 227-230

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Trapping and structural characterisation of a covalent intermediate in vitamin B₆ biosynthesis catalysed by the Pdx1 PLP synthase

M. J. Rodrigues, N. Giri, A. Royant, Y. Zhang, R. Bolton, G. Evans, S. E. Ealick, T. Begley and I. Tews, RSC Chem. Biol., 2022, 3, 227 DOI: 10.1039/D1CB00160D

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