A Single Point Mutation Converts GH84 <i>O</i>-GlcNAc Hydrolases into Phosphorylases: Experimental and Theoretical Evidence

Tezé, David; Coines, Joan; Raich, Lluís; Kalichuk, Valentina; Solleux, Claude; Tellier, Charles; André‐Miral, Corinne; Svensson, Birte; Rovira, Carme

doi:10.1021/jacs.9b09655

Cited by 31 publications

(59 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, GH families 18, 20, 25, 56, 84, 85 and 123 use a substrate‐assisted mechanism, in which an equatorial N ‐acetyl in position C‐2 of the donor acts as the nucleophile [29] . This leads to the formation of a non‐covalent oxazoline‐ or oxazolinium ion‐enzyme intermediate [67] (most likely an oxazolinium ion in the case of GH20 [68] ). Importantly, this type of mechanism predominates in GH‐catalysed reactions involving β‐ d ‐Glc p NAc or β‐ d ‐Gal p NAc, which are highly prevalent in biological systems.…”

Section: Resultsmentioning

confidence: 99%

Rational Enzyme Design without Structural Knowledge: A Sequence‐Based Approach for Efficient Generation of Transglycosylases

Tezé

Zhao

Wiemann

et al. 2021

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

Glycobiology is dogged by the relative scarcity of synthetic, defined oligosaccharides. Enzyme-catalysed glycosylation using glycoside hydrolases is feasible but is hampered by the innate hydrolytic activity of these enzymes. Protein engineering is useful to remedy this, but it usually requires prior structural knowledge of the target enzyme, and/or relies on extensive, time-consuming screening and analysis. Here, a straightforward strategy that involves rational rapid in silico analysis of protein sequences is described. The method pinpoints 6-12 single-mutant candidates to improve transglycosylation yields. Requiring very little prior knowledge of the target enzyme other than its sequence, the method is generic and procures catalysts for the formation of glycosidic bonds involving various d/l-, α/β-pyranosides or furanosides, and exo or endo action. Moreover, mutations validated in one enzyme can be transposed to others, even distantly related enzymes.

show abstract

Section: Resultsmentioning

confidence: 99%

Rational Enzyme Design without Structural Knowledge: A Sequence‐Based Approach for Efficient Generation of Transglycosylases

Tezé

Zhao

Wiemann

et al. 2021

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, GH families 18, 20, 25, 56, 84, 85 and 123 use a substrate-assisted mechanism, in which an equatorial N-acetyl in position C-2 of the donor acts as the nucleophile 47 . This leads to the formation of a non-covalent oxazoline-or oxazolinium ion-enzyme intermediate 48 (oxazolinium ion in the case of GH20 49 ). Importantly, this type of mechanism is predominant in GH-catalyzed reactions involving β-D-GlcNAc or β-D-GalNAc, sugars that are highly prevalent in biological systems.…”

mentioning

confidence: 99%

Rational Enzyme Design Without Structural Knowledge: A Sequence-Based Approach for Efficient Generation of Glycosylation Catalysts

Tezé

Zhao

Wiemann³

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Previous studies on enzyme kinetics and bacterial GH84 homolog structures supported that OGA applied a two‐step, substrate‐assisted mechanism, forming a bicyclic oxazoline intermediate, to hydrolyze O‐GlcNAc modifications (Figure a) . This information has guided the rational design of several potent and selective OGA inhibitors and, more recently, guided the engineering of GH84 enzymes into phosphorylases . As a significant advancement for the field, the structures of human OGA containing the catalytic and stalk domains were reported in 2017 (PDB IDs: 5TKE, 5UHK, and 5M7R) .…”

Section: Structural Insights and Oga Inhibitorsmentioning

confidence: 97%

Molecular Interrogation to Crack the Case of O‐GlcNAc

Estevez

Zhu

Blankenship

et al. 2020

Chemistry A European J

View full text Add to dashboard Cite

The O‐linked β‐N‐acetylglucosamine (O‐GlcNAc) modification, termed O‐GlcNAcylation, is an essential and dynamic post‐translational modification in cells. O‐GlcNAc transferase (OGT) installs this modification on serine and threonine residues, whereas O‐GlcNAcase (OGA) hydrolyzes it. O‐GlcNAc modifications are found on thousands of intracellular proteins involved in diverse biological processes. Dysregulation of O‐GlcNAcylation and O‐GlcNAc cycling enzymes has been detected in many diseases, including cancer, diabetes, cardiovascular and neurodegenerative diseases. Here, recent advances in the development of molecular tools to investigate OGT and OGA functions and substrate recognition are discussed. New chemical approaches to study O‐GlcNAc dynamics and its potential roles in the immune system are also highlighted. It is hoped that this minireview will encourage more research in these areas to advance the understanding of O‐GlcNAc in biology and diseases.

show abstract

A Single Point Mutation Converts GH84 O-GlcNAc Hydrolases into Phosphorylases: Experimental and Theoretical Evidence

Cited by 31 publications

References 37 publications

Rational Enzyme Design without Structural Knowledge: A Sequence‐Based Approach for Efficient Generation of Transglycosylases

Rational Enzyme Design without Structural Knowledge: A Sequence‐Based Approach for Efficient Generation of Transglycosylases

Rational Enzyme Design Without Structural Knowledge: A Sequence-Based Approach for Efficient Generation of Glycosylation Catalysts

Molecular Interrogation to Crack the Case of O‐GlcNAc

Contact Info

Product

Resources

About