Substrate Engagement and Catalytic Mechanisms of N-Acetylglucosaminyltransferase v

John F. Darby; Amelia K. Gilio; Beatriz Piniello; Christian Roth; Elena Blagova; Roderick E. Hubbard; Carme Rovira; Gideon J. Davies; Liang Wu

doi:10.1021/acscatal.0c02222

Substrate Engagement and Catalytic Mechanisms of N-Acetylglucosaminyltransferase v

John F. Darby, Amelia K. Gilio, Beatriz Piniello, Christian Roth, Elena Blagova, Roderick E. Hubbard, Carme Rovira, Gideon J. Davies^*, Liang Wu

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

α-Mannoside β-1,6-N-acetylglucosaminyltransferase V (MGAT5) is a mammalian glycosyltransferase involved in complex N-glycan formation, which strongly drives cancer when overexpressed. Despite intense interest, the catalytic mechanism of MGAT5 is not known in detail, precluding therapeutic exploitation. We solved structures of MGAT5 complexed to glycosyl donor and acceptor ligands, revealing an unforeseen role for donor-induced loop rearrangements in controlling acceptor substrate engagement. QM/MM metadynamics simulations of MGAT5 catalysis highlight the key assisting role of Glu297 and reveal considerable conformational distortions imposed upon the glycosyl donor during transfer. Detailed mechanistic characterization of MGAT5 will aid inhibitor development to correct cancer-associated N-glycosylation.

Original language	English
Pages (from-to)	8590-8596
Number of pages	7
Journal	ACS Catalysis
Volume	10
Issue number	15
Early online date	16 Jul 2020
DOIs	https://doi.org/10.1021/acscatal.0c02222
Publication status	Published - 7 Aug 2020

Bibliographical note

Keywords

carbohydrates
enzymes
glycosyltransferases
N-glycosylation
quantum mechanics/molecular mechanics

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10.1021/acscatal.0c02222

Substrate Engagement and Catalytic Mechanisms of N‑Acetylglucosaminyltransferase V
© 2020 American Chemical Society
Final published version, 4.78 MBLicence: CC BY

Cite this

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title = "Substrate Engagement and Catalytic Mechanisms of N-Acetylglucosaminyltransferase v",

abstract = "α-Mannoside β-1,6-N-acetylglucosaminyltransferase V (MGAT5) is a mammalian glycosyltransferase involved in complex N-glycan formation, which strongly drives cancer when overexpressed. Despite intense interest, the catalytic mechanism of MGAT5 is not known in detail, precluding therapeutic exploitation. We solved structures of MGAT5 complexed to glycosyl donor and acceptor ligands, revealing an unforeseen role for donor-induced loop rearrangements in controlling acceptor substrate engagement. QM/MM metadynamics simulations of MGAT5 catalysis highlight the key assisting role of Glu297 and reveal considerable conformational distortions imposed upon the glycosyl donor during transfer. Detailed mechanistic characterization of MGAT5 will aid inhibitor development to correct cancer-associated N-glycosylation.",

keywords = "carbohydrates, enzymes, glycosyltransferases, N-glycosylation, quantum mechanics/molecular mechanics",

author = "Darby, {John F.} and Gilio, {Amelia K.} and Beatriz Piniello and Christian Roth and Elena Blagova and Hubbard, {Roderick E.} and Carme Rovira and Davies, {Gideon J.} and Liang Wu",

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AU - Darby, John F.

AU - Gilio, Amelia K.

AU - Piniello, Beatriz

AU - Roth, Christian

AU - Blagova, Elena

AU - Hubbard, Roderick E.

AU - Rovira, Carme

AU - Davies, Gideon J.

AU - Wu, Liang

PY - 2020/8/7

Y1 - 2020/8/7

N2 - α-Mannoside β-1,6-N-acetylglucosaminyltransferase V (MGAT5) is a mammalian glycosyltransferase involved in complex N-glycan formation, which strongly drives cancer when overexpressed. Despite intense interest, the catalytic mechanism of MGAT5 is not known in detail, precluding therapeutic exploitation. We solved structures of MGAT5 complexed to glycosyl donor and acceptor ligands, revealing an unforeseen role for donor-induced loop rearrangements in controlling acceptor substrate engagement. QM/MM metadynamics simulations of MGAT5 catalysis highlight the key assisting role of Glu297 and reveal considerable conformational distortions imposed upon the glycosyl donor during transfer. Detailed mechanistic characterization of MGAT5 will aid inhibitor development to correct cancer-associated N-glycosylation.

AB - α-Mannoside β-1,6-N-acetylglucosaminyltransferase V (MGAT5) is a mammalian glycosyltransferase involved in complex N-glycan formation, which strongly drives cancer when overexpressed. Despite intense interest, the catalytic mechanism of MGAT5 is not known in detail, precluding therapeutic exploitation. We solved structures of MGAT5 complexed to glycosyl donor and acceptor ligands, revealing an unforeseen role for donor-induced loop rearrangements in controlling acceptor substrate engagement. QM/MM metadynamics simulations of MGAT5 catalysis highlight the key assisting role of Glu297 and reveal considerable conformational distortions imposed upon the glycosyl donor during transfer. Detailed mechanistic characterization of MGAT5 will aid inhibitor development to correct cancer-associated N-glycosylation.

KW - carbohydrates

KW - enzymes

KW - glycosyltransferases

KW - N-glycosylation

KW - quantum mechanics/molecular mechanics

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JO - ACS Catalysis

JF - ACS Catalysis

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Substrate Engagement and Catalytic Mechanisms of N-Acetylglucosaminyltransferase v

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

RS Research Professorship

Fragment-based activation and modification of industrial enzymes

Glycosylation: Programmes for Observation, Inhibition & Structure-based Exploitation of key carbohydrate-active enzymes

Cite this

Substrate Engagement and Catalytic Mechanisms of N-Acetylglucosaminyltransferase v

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Projects

RS Research Professorship

Fragment-based activation and modification of industrial enzymes

Glycosylation: Programmes for Observation, Inhibition & Structure-based Exploitation of key carbohydrate-active enzymes

Cite this