DeciphGYG · Deciphering the Molecular Mechanism of an Enzymatic Machinery for Glycogen Biosynthesis
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2022-05-02 → 2024-05-01
- EU contribution
- €172,932
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Deciphering the Molecular Mechanism of an Enzymatic Machinery for Glycogen Biosynthesis
The project “Deciphering the Molecular Mechanism of an Enzymatic Machinery for Glycogen Biosynthesis” focuses on addressing the mechanism of the enzymatic machinery of human glycogenin 1 for glycogen biosynthesis. How is the substrate-induced conformational change coupled with the chemical reactions? What is the mechanism of chemical reactions at different lengths of the glucose chain as an acceptor? How does the mutation T83M lead to enzymatically inactive, causing glycogen storage diseases? A new protocol is planned to be developed to answer these questions, which is transferable to other glycosyltransferases. The research idea of this work was that many mutations of enzymes could lead to diseases, and the mechanism behind the mutations is still not well understood in most cases due to limited capacity and technical caveats of the available research tools. We aimed to combine both multiscale computational modeling methods and experimental tools to uncover the deep mechanism. Such a combined approach has not yet been very common in this field. Working on this project might increase understanding of how enzymes function and how mutations lead to the malfunction of enzymes. Deeper knowledge will facilitate drug design against mutation-associated diseases with more confidence, which will improve the biotechnology development in the pharmaceutical industry. The objectives of this MSCA project are (1) to identify the Michaelis complexes of hGYG1 with different lengths of sugar chains as acceptors and study their recognition. (2) to address the catalytic mechanism of glucosyl transfer within hGYG1 (3) to investigate the effects of mutation T83M on the catalysis and dynamics of hGYG1. How a single mutation on the lid could lead to an inactive hGYG1, causing glycogen storage disease.
Data: CORDIS, © European Union
Project objective
The biosynthesis of glycogen - glycogenesis - represents a key glucose (and hence energy) storage process across a wide range of organisms. Human glycogenin 1 (hGYG1) is one of the two primary enzymes that initiates the biosynthesis of glycogen, our energy reservoir. It polymerizes a maltosaccharide chain covalently attached to an enzyme residue, Y195, via a stepwise glycosylation reaction. In the reaction cycle, a dynamic conformational switch between ground and active states induced by one of the enzyme substrates, the sugar donor UDP-glucose, was predicted by structural studies, including a stretch of a critical loop containing Y195, the acceptor arm, and a major movement of a 32-residue lid covering the active site. The T83M mutation, which causes glycogen storage disease (GSD) type XV, makes the enzyme catalytically inactive. The scientific aim of DeciphGYG is to design a new protocol combining a series of both computational techniques (MD, QM/MM MetaD, HREX and BE-MetaD) and experimental results (NMR and crystallography) to unveil the mechanisms of glycogen biosynthesis, including both the glycosylation reaction as well as its coupling with the conformational changes induced by binding of the UDP-glucose donor and acceptor substrates. The experienced researcher (ER), Qinghua Liao will carry out the project in the University of Barcelona under the supervision of Prof. Carme Rovira, who has extensive experience in computational modeling of carbohydrate-active enzymes. Our goal will be shaping the understanding of hGYG1 action fully on glycogen biosynthesis, facilitating drug design against GSD type XV. Moreover, the new protocol will be transferable to other enzymes of interest in glycobiology. Altogether, the DeciphGYG project will allow the ER with a highly competitive multidisciplinary profile by complementing his previous acquired skills, placing him at a strong position to start his career as an independent and innovative principal investigator.
Original text from CORDIS.
Participants
- UNIVERSITAT DE BARCELONA · BarcelonaCoordinatorSpain
Links
Data: CORDIS, © European Union