Analysis of Protein O‐GlcNAcylation by Mass Spectrometry

O‐linked β‐D‐N‐acetyl glucosamine (O‐GlcNAc) addition (O‐GlcNAcylation), a post‐translational modification of serine/threonine residues of proteins, is involved in diverse cellular metabolic and signaling pathways. Aberrant O‐GlcNAcylation underlies the initiation and progression of multiple chronic diseases including diabetes, cancer, and neurodegenerative diseases. Numerous methods have been developed for the analysis of protein O‐GlcNAcylation, but instead of discussing the classical biochemical techniques, this unit covers O‐GlcNAc characterization by combining several enrichment methods and mass spectrometry detection techniques [including collision‐induced dissociation (CID), higher energy collision dissociation (HCD), and electron transfer dissociation (ETD) mass spectrometry]. © 2017 by John Wiley & Sons, Inc.

[1]  J. Shabanowitz,et al.  Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  G. Hart Three Decades of Research on O-GlcNAcylation – A Major Nutrient Sensor That Regulates Signaling, Transcription and Cellular Metabolism , 2014, Front. Endocrinol..

[3]  Robert J Chalkley,et al.  Identification of protein O-GlcNAcylation sites using electron transfer dissociation mass spectrometry on native peptides , 2009, Proceedings of the National Academy of Sciences.

[4]  Susana Comte-Walters,et al.  Identification of O-Linked N-Acetylglucosamine (O-GlcNAc)-modified Osteoblast Proteins by Electron Transfer Dissociation Tandem Mass Spectrometry Reveals Proteins Critical for Bone Formation* , 2013, Molecular & Cellular Proteomics.

[5]  Matthew E Monroe,et al.  Tandem mass spectrometry identifies many mouse brain O-GlcNAcylated proteins including EGF domain-specific O-GlcNAc transferase targets , 2012, Proceedings of the National Academy of Sciences.

[6]  G. Hart,et al.  Site-Specific GlcNAcylation of Human Erythrocyte Proteins , 2009, Diabetes.

[7]  S. Ficarro,et al.  Exploring the O-GlcNAc proteome: direct identification of O-GlcNAc-modified proteins from the brain. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Larsen,et al.  Purification and identification of O-GlcNAc-modified peptides using phosphate-based alkyne CLICK chemistry in combination with titanium dioxide chromatography and mass spectrometry. , 2011, Journal of proteome research.

[9]  R. Cole,et al.  Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle , 2015, Diabetes.

[10]  Lance Wells,et al.  Combining high-energy C-trap dissociation and electron transfer dissociation for protein O-GlcNAc modification site assignment. , 2011, Journal of proteome research.

[11]  B. Kuster,et al.  Proteome Wide Purification and Identification of O‑glcnac-modified Proteins Using Click Chemistry and Mass Spectrometry , 2022 .

[12]  Scott B Ficarro,et al.  Probing the dynamics of O-GlcNAc glycosylation in the brain using quantitative proteomics. , 2007, Nature chemical biology.

[13]  R. Chalkley,et al.  Chapter 15 O-GlcNAc Proteomics: Mass Spectrometric Analysis of O-GlcNAc Modifications on Proteins , 2008 .

[14]  Animesh Nandi,et al.  Global identification of O-GlcNAc-modified proteins. , 2006, Analytical chemistry.

[15]  J. Hanover,et al.  JCB_201501101 1..12 , 2015 .

[16]  Jonathan C Trinidad,et al.  O-Linked N-Acetylglucosamine Proteomics of Postsynaptic Density Preparations Using Lectin Weak Affinity Chromatography and Mass Spectrometry*S , 2006, Molecular & Cellular Proteomics.

[17]  G. Hart,et al.  Topography and polypeptide distribution of terminal N-acetylglucosamine residues on the surfaces of intact lymphocytes. Evidence for O-linked GlcNAc. , 1984, The Journal of biological chemistry.

[18]  M. Wolfert,et al.  Glycopeptide-specific monoclonal antibodies suggest new roles for O-GlcNAc. , 2010, Nature chemical biology.

[19]  Junfeng Ma,et al.  O-GlcNAcomic Profiling Identifies Widespread O-Linked β-N-Acetylglucosamine Modification (O-GlcNAcylation) in Oxidative Phosphorylation System Regulating Cardiac Mitochondrial Function*♦ , 2015, The Journal of Biological Chemistry.

[20]  B K Hayes,et al.  Selective detection and site-analysis of O-GlcNAc-modified glycopeptides by beta-elimination and tandem electrospray mass spectrometry. , 1996, Analytical biochemistry.

[21]  Junfeng Ma,et al.  O-GlcNAc profiling: from proteins to proteomes , 2014, Clinical Proteomics.

[22]  A. Pandey,et al.  The dynamic stress-induced “O-GlcNAc-ome” highlights functions for O-GlcNAc in regulating DNA damage/repair and other cellular pathways , 2011, Amino Acids.

[23]  G. Hart,et al.  The subcellular distribution of terminal N-acetylglucosamine moieties. Localization of a novel protein-saccharide linkage, O-linked GlcNAc. , 1986, The Journal of biological chemistry.

[24]  Lance Wells,et al.  Mapping Sites of O-GlcNAc Modification Using Affinity Tags for Serine and Threonine Post-translational Modifications* , 2002, Molecular & Cellular Proteomics.

[25]  Lance Wells,et al.  Quantitative analysis of both protein expression and serine / threonine post‐translational modifications through stable isotope labeling with dithiothreitol , 2005, Proteomics.

[26]  Junfeng Ma,et al.  Protein O-GlcNAcylation in diabetes and diabetic complications , 2013, Expert review of proteomics.

[27]  R. Cole,et al.  Detection and Analysis of Proteins Modified by O‐Linked N‐Acetylglucosamine , 2001, Current protocols in protein science.

[28]  J. Shabanowitz,et al.  Extensive Crosstalk Between O-GlcNAcylation and Phosphorylation Regulates Cytokinesis , 2010, Science Signaling.

[29]  A. Pandey,et al.  Dynamic Interplay between O-Linked N-Acetylglucosaminylation and Glycogen Synthase Kinase-3-dependent Phosphorylation* , 2007, Molecular & Cellular Proteomics.

[30]  Carolyn R Bertozzi,et al.  A chemical approach for identifying O-GlcNAc-modified proteins in cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Junfeng Ma,et al.  Mass Spectrometry-Based Quantitative O-GlcNAcomic Analysis. , 2016, Methods in molecular biology.

[32]  A. Burlingame,et al.  Global Identification and Characterization of Both O-GlcNAcylation and Phosphorylation at the Murine Synapse* , 2012, Molecular & Cellular Proteomics.

[33]  J. Shabanowitz,et al.  Enrichment and Site Mapping of O-Linked N-Acetylglucosamine by a Combination of Chemical/Enzymatic Tagging, Photochemical Cleavage, and Electron Transfer Dissociation Mass Spectrometry* , 2009, Molecular & Cellular Proteomics.

[34]  G. Hart,et al.  Glycomic Approaches to Study GlcNAcylation: Protein Identification, Site-mapping, and Site-specific O-GlcNAc Quantitation , 2008, Clinical Proteomics.

[35]  Jennifer J. Kohler,et al.  Metabolic cross-talk allows labeling of O-linked β-N-acetylglucosamine-modified proteins via the N-acetylgalactosamine salvage pathway , 2011, Proceedings of the National Academy of Sciences.