Methods for Studying Site-Specific O-GlcNAc Modifications: Successes, Limitations, and Important Future Goals

Moon, Stuart P.; Javed, Afraah; Hard, Eldon R.; Pratt, Matthew R.

doi:10.1021/jacsau.1c00455

Cited by 15 publications

(14 citation statements)

References 92 publications

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“…[10,11] Moreover, the advent of bioorthogonal chemistry enabled labeling of saccharide of interests in a complicated mixture of biomolecules including in cell and in vivo environment to interrogate their roles in living systems. [12][13][14][15][16][17] Fundamental challenges of selective labeling of saccharides stem from their chemical diversity with highly oxygen-rich nature. The polyol structure not only hinders selective labeling of a target functional group, but the presence of multiple hydrophilic groups also results in their poor solubility in organic solvents.…”

Section: Introductionmentioning

confidence: 99%

Phosphine-Mediated Three-Component Bioconjugation of Amino- and Azidosaccharides in Ionic Liquids

Ohata

Hall

Uzoewulu

et al. 2022

Preprint

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Bioconjugation of carbohydrates has advanced the modern chemical biology and medical science research by incorporation of desired functionalities to the oxygen-rich biomolecules. However, the labeling process or selective chemical reaction of carbohydrates have been a challenging task because of their chemical, functional, and structural diversities, and no single chemical modification tool can be universally applicable to all the target substrates in different environments. In this report, we have developed a bioconjugation strategy for labeling of carbohydrate derivatives through a phosphine-mediated three-component coupling reaction in an ionic liquid medium. The multiple characterization methods identified a urea group as the reaction product of the phosphine-mediated coupling through the installation of carbonyl group from carbon dioxide in air and loss of nitrogen gas from the azide group. We have developed purification protocols to facilitate the cleanup and analysis of ionic liquid-based bioconjugation processes, which can be used for a diverse set of carbohydrate derivatives. The phosphine-mediated urea-forming reaction was applied to a variety of amine- and azide-containing carbohydrates such as antibiotics, anti-tumor agent, and polysaccharide with corresponding azide- and amine-based reagent, respectively. The ionic liquid-based bioconjugation was amenable for the chemical modification of the mammalian cell lysate in an azide-dependent manner. Thus, the present report represents not only the advancement of the nonaqueous bioconjugation method for carbohydrate derivatives from the reaction development perspectives, but also their practical utility for creation of carbohydrate conjugates as well as a study tool of the biomolecule through the bioorthogonal chemistry-like reactivity.

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Section: Introductionmentioning

confidence: 99%

Phosphine-Mediated Three-Component Bioconjugation of Amino- and Azidosaccharides in Ionic Liquids

Ohata

Hall

Uzoewulu

et al. 2022

Preprint

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“…Several strategies of chemical synthesis were implemented to prevent aggregation into β-sheet structures and improve solubility of difficult sequences ( Butterfield et al, 2012 ), but these considerations are beyond the scope of this review. NCL, initially developed by Kent and co-workers, uses chemoselective reactions between the α-carboxyl and the α-amino groups of two unprotected peptides to form a native peptide bond ( Muir and Kent, 1993 ; Dawson et al, 1994 ; Hackenberger and Schwarzer, 2008 ; Agouridas et al, 2019 ; Moon et al, 2022 ). This strategy allows introducing selective and quantitative modifications of amino acids without altering any usual peptide bond.…”

Section: Deciphering the Posttranslational Modification Codes Of Amyl...mentioning

confidence: 99%

“…The former approach uses the manipulation of the UDP-GlcNAc metabolic pathway and the OGT plasticity to a variety of UDP-GlcNAc derivatives (e.g. peracetylated N-azidoacetylglucosamine) to directly incorporate chemical reactive group in the GlcNAc moiety of glycosylated proteins ( Moon et al, 2022 ; Saha et al, 2021 ). The latter approach achieved better efficiency by using a mutant of GalT (GalT-Y289L) tolerant to UDP-GalNAc derivatives, such UDP-GalNAz containing an azide group, as a substrate donor for glycosidic linkage to GlcNAc ( Figure 7 ) ( Khidekel et al, 2003 ).…”

Section: Deciphering the Posttranslational Modification Codes Of Amyl...mentioning

confidence: 99%

Deciphering the Structure and Formation of Amyloids in Neurodegenerative Diseases With Chemical Biology Tools

et al. 2022

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Protein aggregation into highly ordered, regularly repeated cross-β sheet structures called amyloid fibrils is closely associated to human disorders such as neurodegenerative diseases including Alzheimer’s and Parkinson’s diseases, or systemic diseases like type II diabetes. Yet, in some cases, such as the HET-s prion, amyloids have biological functions. High-resolution structures of amyloids fibrils from cryo-electron microscopy have very recently highlighted their ultrastructural organization and polymorphisms. However, the molecular mechanisms and the role of co-factors (posttranslational modifications, non-proteinaceous components and other proteins) acting on the fibril formation are still poorly understood. Whether amyloid fibrils play a toxic or protective role in the pathogenesis of neurodegenerative diseases remains to be elucidated. Furthermore, such aberrant protein-protein interactions challenge the search of small-molecule drugs or immunotherapy approaches targeting amyloid formation. In this review, we describe how chemical biology tools contribute to new insights on the mode of action of amyloidogenic proteins and peptides, defining their structural signature and aggregation pathways by capturing their molecular details and conformational heterogeneity. Challenging the imagination of scientists, this constantly expanding field provides crucial tools to unravel mechanistic detail of amyloid formation such as semisynthetic proteins and small-molecule sensors of conformational changes and/or aggregation. Protein engineering methods and bioorthogonal chemistry for the introduction of protein chemical modifications are additional fruitful strategies to tackle the challenge of understanding amyloid formation.

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“…Inducing constitutively active mutants at identified PTM sites is a powerful tool for phosphorylation events; however, the O-GlcNAc modification does not contain unique chemical moieties that can be mimicked with an amino acid substitution. In contrast, amino acid substitutions to permanently remove identified O-GlcNAc sites have been employed with success to explore cell signaling mechanisms ( Housley et al, 2008 ; Myers et al, 2016 ; Wu et al, 2016 ; Kim et al, 2020 ) (reviewed in ( Moon et al, 2022 )).…”

Section: Introductionmentioning

confidence: 99%

Mapping the O-GlcNAc Modified Proteome: Applications for Health and Disease

Burt

Alghusen

Ephrame

et al. 2022

Front. Mol. Biosci.

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O-GlcNAc is a pleotropic, enigmatic post-translational modification (PTM). This PTM modifies thousands of proteins differentially across tissue types and regulates diverse cellular signaling processes. O-GlcNAc is implicated in numerous diseases, and the advent of O-GlcNAc perturbation as a novel class of therapeutic underscores the importance of identifying and quantifying the O-GlcNAc modified proteome. Here, we review recent advances in mass spectrometry-based proteomics that will be critical in elucidating the role of this unique glycosylation system in health and disease.

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Methods for Studying Site-Specific O-GlcNAc Modifications: Successes, Limitations, and Important Future Goals

Cited by 15 publications

References 92 publications

Phosphine-Mediated Three-Component Bioconjugation of Amino- and Azidosaccharides in Ionic Liquids

Phosphine-Mediated Three-Component Bioconjugation of Amino- and Azidosaccharides in Ionic Liquids

Deciphering the Structure and Formation of Amyloids in Neurodegenerative Diseases With Chemical Biology Tools

Mapping the O-GlcNAc Modified Proteome: Applications for Health and Disease

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