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由Pdx1磷酸吡哆醛合酶催化的维生素B生物合成中共价中间体的捕获与结构表征

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

作者信息

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

机构信息

Biological Sciences, Institute for Life Sciences, University of Southampton Southampton SO17 1BJ UK

Diamond Light Source, Harwell Science and Innovation Campus Didcot OX11 0DE UK.

出版信息

RSC Chem Biol. 2021 Oct 25;3(2):227-230. doi: 10.1039/d1cb00160d. eCollection 2022 Feb 9.

DOI:10.1039/d1cb00160d
PMID:35360887
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8827014/
Abstract

The Pdx1 enzyme catalyses condensation of two carbohydrates and ammonia to form pyridoxal 5-phosphate (PLP) an imine relay mechanism of carbonyl intermediates. The I 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.

摘要

Pdx1 酶催化两种碳水化合物与氨缩合形成磷酸吡哆醛（PLP），这是一种通过羰基中间体的亚胺中继机制。本文利用结构、紫外可见吸收光谱和质谱分析对 I 中间体进行了表征，该中间体解释了立体选择性去质子化以及随后底物辅助的磷酸消除反应，这是 PLP 生物合成的核心。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/b8331a06c379/d1cb00160d-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/e80824a9f43a/d1cb00160d-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/69048941ee0b/d1cb00160d-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/6eb38fb709ec/d1cb00160d-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/f96f7e347482/d1cb00160d-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/b8331a06c379/d1cb00160d-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/e80824a9f43a/d1cb00160d-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/69048941ee0b/d1cb00160d-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/6eb38fb709ec/d1cb00160d-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/f96f7e347482/d1cb00160d-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc95/8827014/b8331a06c379/d1cb00160d-f5.jpg

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Acta Crystallogr D Struct Biol. 2019 Apr 1;75(Pt 4):400-415. doi: 10.1107/S2059798319002912. Epub 2019 Apr 4.
2
Lysine relay mechanism coordinates intermediate transfer in vitamin B6 biosynthesis.赖氨酸中继机制协调维生素B6生物合成中的中间体转移。
Nat Chem Biol. 2017 Mar;13(3):290-294. doi: 10.1038/nchembio.2273. Epub 2017 Jan 16.
3
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Proc Natl Acad Sci U S A. 2016 Oct 4;113(40):E5821-E5829. doi: 10.1073/pnas.1608125113. Epub 2016 Sep 19.
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Challenges and solutions for the analysis of in situ, in crystallo micro-spectrophotometric data.原位、晶体中显微分光光度数据的分析挑战与解决方案
Acta Crystallogr D Biol Crystallogr. 2015 Jan 1;71(Pt 1):27-35. doi: 10.1107/S1399004714015107.
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In crystallo optical spectroscopy (icOS) as a complementary tool on the macromolecular crystallography beamlines of the ESRF.作为欧洲同步辐射装置（ESRF）大分子晶体学光束线上的一种辅助工具的晶体光学光谱学（icOS）。
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