Literature DB >> 34004049

Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine.

Kamau Fahie1, Bhargavi Narayanan1, Fiddia Zahra1, Russell Reeves1,2, Steve M Fernandes1, Gerald W Hart3, Natasha E Zachara1.   

Abstract

O-GlcNAc is a common post-translational modification of nuclear, mitochondrial, and cytoplasmic proteins that regulates normal physiology and the cell stress response. Dysregulation of O-GlcNAc cycling is implicated in the etiology of type II diabetes, heart failure, hypertension, and Alzheimer's disease, as well as cardioprotection. These protocols cover simple and comprehensive techniques for detecting proteins modified by O-GlcNAc and studying the enzymes that add or remove O-GlcNAc.
© 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Increasing the stoichiometry of O-GlcNAc on proteins before analysis Basic Protocol 2: Detection of proteins modified by O-GlcNAc using antibodies Basic Protocol 3: Detection of proteins modified by O-GlcNAc using the lectin sWGA Support Protocol 1: Control for O-linked glycosylation Basic Protocol 4: Detection and enrichment of proteins using WGA-agarose Support Protocol 2: Digestion of proteins with hexosaminidase Alternate Protocol: Detection of proteins modified by O-GlcNAc using galactosyltransferase Support Protocol 3: Autogalactosylation of galactosyltransferase Support Protocol 4: Assay of galactosyltransferase activity Basic Protocol 5: Characterization of labeled glycans by β-elimination and chromatography Basic Protocol 6: Detection of O-GlcNAc in 96-well plates Basic Protocol 7: Assay for OGT activity Support Protocol 5: Desalting of O-GlcNAc transferase Basic Protocol 8: Assay for O-GlcNAcase activity. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC.

Entities:  

Keywords:  O-GlcNAc; O-linked; analysis; detection; galactosyltransferase; glycosylation; signal transduction

Mesh:

Substances:

Year:  2021        PMID: 34004049      PMCID: PMC8862748          DOI: 10.1002/cpz1.129

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  99 in total

1.  Quantitative conversion of mucin-type sugar chains to radioactive oligosaccharides.

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2.  A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo.

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5.  Dynamic O-glycosylation of nuclear and cytosolic proteins: cloning and characterization of a neutral, cytosolic beta-N-acetylglucosaminidase from human brain.

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6.  Responsiveness of the state of O-linked N-acetylglucosamine modification of nuclear pore protein p62 to the extracellular glucose concentration.

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Review 7.  Cross talk between O-GlcNAcylation and phosphorylation: roles in signaling, transcription, and chronic disease.

Authors:  Gerald W Hart; Chad Slawson; Genaro Ramirez-Correa; Olof Lagerlof
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Authors:  H H Freeze
Journal:  Curr Protoc Protein Sci       Date:  2001-05

9.  Unique hexosaminidase reduces metabolic survival signal and sensitizes cardiac myocytes to hypoxia/reoxygenation injury.

Authors:  Gladys A Ngoh; Heberty T Facundo; Tariq Hamid; Wolfgang Dillmann; Natasha E Zachara; Steven P Jones
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10.  Cytokeratin peptide SFGSGFGGGY mimics N-acetyl-beta-D-glucosamine in reaction with antibodies and lectins, and induces in vivo anti-carbohydrate antibody response.

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Journal:  J Immunol       Date:  1994-12-15       Impact factor: 5.422

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  2 in total

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2.  Altered protein O-GlcNAcylation in placentas from mothers with diabetes causes aberrant endocytosis in placental trophoblast cells.

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  2 in total

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