O-GlcNAc profiling: from proteins to proteomes

O-linked β-D-N-acetylglucosamine (O-GlcNAc) modification (O-GlcNAcylation) onto serine and threonine residues of proteins is an important post-translational modification (PTM), which is involved in many crucial biological processes including transcription, translation, proteasomal degradation, and signal transduction. Aberrant protein O-GlcNAcylation is directly linked to the pathological progression of chronic diseases including diabetes, cancer, and neurodegenerative disorders. Identification, site mapping, and quantification of O-GlcNAc proteins are a prerequisite to decipher their functions. In this review, we mainly focus on technological developments regarding O-GlcNAc protein profiling. Specifically, on one hand, we show how these techniques are being used for the comprehensive characterization of certain targeted proteins in which biologists are most interested. On the other hand, we present several newly developed approaches for O-GlcNAcomic profiling as well as how they provide us with a systems perspective to crosstalk amongst different PTMs and complicated biological events. Promising technical trends are also highlighted to evoke more efforts by diverse laboratories, which would further expand our understanding of the physiological and pathological roles of protein O-GlcNAcylation in chronic diseases.

[1]  R. Aebersold,et al.  Mass spectrometry-based proteomics and network biology. , 2012, Annual review of biochemistry.

[2]  Yi E. Sun,et al.  Quantitation of O-Glycosylation Stoichiometry and Dynamics using Resolvable Mass Tags , 2010, Nature chemical biology.

[3]  G. Hart,et al.  Purification and characterization of an O-GlcNAc selective N-acetyl-beta-D-glucosaminidase from rat spleen cytosol. , 1994, The Journal of biological chemistry.

[4]  T. Lefebvre,et al.  Characterization of O-GlcNAc cycling and proteomic identification of differentially O-GlcNAcylated proteins during G1/S transition. , 2012, Biochimica et biophysica acta.

[5]  M. Karas,et al.  Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. , 1988, Analytical chemistry.

[6]  S. Marshall,et al.  Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system. Role of hexosamine biosynthesis in the induction of insulin resistance. , 1991, The Journal of biological chemistry.

[7]  T. Isono.  O-GlcNAc-Specific Antibody CTD110.6 Cross-Reacts with N-GlcNAc2-Modified Proteins Induced under Glucose Deprivation , 2011, PloS one.

[8]  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.

[9]  Reinout Raijmakers,et al.  Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics , 2009, Nature Protocols.

[10]  S. Jacobsen,et al.  SPINDLY, a tetratricopeptide repeat protein involved in gibberellin signal transduction in Arabidopsis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

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

[12]  G. Hart,et al.  Enzymatic addition of O-GlcNAc to nuclear and cytoplasmic proteins. Identification of a uridine diphospho-N-acetylglucosamine:peptide beta-N-acetylglucosaminyltransferase. , 1990, The Journal of biological chemistry.

[13]  G. Hart,et al.  Characterization of a mouse monoclonal antibody specific for O-linked N-acetylglucosamine. , 2001, Analytical biochemistry.

[14]  David R M Graham,et al.  Two‐dimensional gel‐based approaches for the assessment of N‐Linked and O‐GlcNAc glycosylation in human and simian immunodeficiency viruses , 2008, Proteomics.

[15]  C. Srisomsap,et al.  Proteomic analysis and abrogated expression of O‐GlcNAcylated proteins associated with primary breast cancer , 2013, Proteomics.

[16]  K. Vosseller,et al.  O-GlcNAc in cancer biology , 2013, Amino Acids.

[17]  M. Mann,et al.  SILAC Mouse for Quantitative Proteomics Uncovers Kindlin-3 as an Essential Factor for Red Blood Cell Function , 2008, Cell.

[18]  J. Hirabayashi.  Lectin-based structural glycomics: Glycoproteomics and glycan profiling , 2004, Glycoconjugate Journal.

[19]  T. Lefebvre,et al.  O-GlcNAcylation: A New Cancer Hallmark? , 2013, Front. Endocrinol..

[20]  T. Rabilloud,et al.  Two-dimensional gel electrophoresis in proteomics: Past, present and future. , 2010, Journal of proteomics.

[21]  M. Mann,et al.  A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC) , 2006, Nature Protocols.

[22]  G. Hart,et al.  Regulation of CK2 by Phosphorylation and O-GlcNAcylation Revealed by Semisynthesis , 2011, Nature chemical biology.

[23]  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.

[24]  B. Ueberheide,et al.  The utility of ETD mass spectrometry in proteomic analysis. , 2006, Biochimica et biophysica acta.

[25]  Akhilesh Pandey,et al.  Quantitative proteomics using stable isotope labeling with amino acids in cell culture , 2008, Nature Protocols.

[26]  Linda C Hsieh-Wilson,et al.  A chemoenzymatic approach toward the rapid and sensitive detection of O-GlcNAc posttranslational modifications. , 2003, Journal of the American Chemical Society.

[27]  G. Hart,et al.  Regulation of Calcium/Calmodulin-dependent Kinase IV by O-GlcNAc Modification* , 2009, The Journal of Biological Chemistry.

[28]  J. Hirabayashi,et al.  Comparative analysis by frontal affinity chromatography of oligosaccharide specificity of GlcNAc-binding lectins, Griffonia simplicifolia lectin-II (GSL-II) and Boletopsis leucomelas lectin (BLL). , 2006, Journal of biochemistry.

[29]  G. Hart,et al.  Specific isolation of O-linked N-acetylglucosamine glycopeptides from complex mixtures. , 1995, Analytical biochemistry.

[30]  J. Shabanowitz,et al.  O-Linked β-N-Acetylglucosamine (O-GlcNAc) Regulates Emerin Binding to Barrier to Autointegration Factor (BAF) in a Chromatin- and Lamin B-enriched “Niche”* , 2013, The Journal of Biological Chemistry.

[31]  Jeffrey W. Smith,et al.  Mass Spectrometry-Based Label-Free Quantitative Proteomics , 2009, Journal of biomedicine & biotechnology.

[32]  T. Issad,et al.  O-GlcNAc modification, insulin signaling and diabetic complications. , 2010, Diabetes & metabolism.

[33]  G. Hart,et al.  The intersections between O-GlcNAcylation and phosphorylation: implications for multiple signaling pathways , 2010, Journal of Cell Science.

[34]  Sanjeeva Srivastava,et al.  Proteomic databases and tools to decipher post-translational modifications. , 2011, Journal of proteomics.

[35]  T. Lam,et al.  Posttranslational modifications on protein kinase c isozymes. Effects of epinephrine and phorbol esters. , 2008, Biochimica et biophysica acta.

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

[37]  M. Mann,et al.  Electrospray ionization for mass spectrometry of large biomolecules. , 1989, Science.

[38]  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.

[39]  Michael Barber,et al.  Fast atom bombardment of solids (F.A.B.): a new ion source for mass spectrometry , 1981 .

[40]  R. Chalkley,et al.  Identification of GlcNAcylation sites of peptides and α-crystallin using Q-TOF mass spectrometry , 2001 .

[41]  G. Hart,et al.  O-Linked GlcNAc Modification of Cardiac Myofilament Proteins: A Novel Regulator of Myocardial Contractile Function , 2008, Circulation research.

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

[43]  D. Baltimore,et al.  Activation of the Transcriptional Function of the NF-κB Protein c-Rel by O-GlcNAc Glycosylation , 2013, Science Signaling.

[44]  M. Taussig,et al.  Protein microarrays: high-throughput tools for proteomics , 2009, Expert review of proteomics.

[45]  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.

[46]  N. Greenspan,et al.  Cytologic assessment of nuclear and cytoplasmic O-linked N-acetylglucosamine distribution by using anti-streptococcal monoclonal antibodies. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Søren Brunak,et al.  Prediction of Glycosylation Across the Human Proteome and the Correlation to Protein Function , 2001, Pacific Symposium on Biocomputing.

[48]  S. Trauger,et al.  Increased Enzymatic O-GlcNAcylation of Mitochondrial Proteins Impairs Mitochondrial Function in Cardiac Myocytes Exposed to High Glucose* , 2009, Journal of Biological Chemistry.

[49]  J. Hanover,et al.  Mitochondrial and nucleocytoplasmic isoforms of O-linked GlcNAc transferase encoded by a single mammalian gene. , 2003, Archives of biochemistry and biophysics.

[50]  J. Hanover,et al.  O-GlcNAc cycling: implications for neurodegenerative disorders. , 2009, The international journal of biochemistry & cell biology.

[51]  J. Michalski,et al.  Mapping of O‐linked β‐N‐acetylglucosamine modification sites in key contractile proteins of rat skeletal muscle , 2009, Proteomics.

[52]  A. Varki,et al.  Evolution of Glycan Diversity , 2009 .

[53]  G. Hart,et al.  O-linked beta-N-acetylglucosamine (O-GlcNAc): Extensive crosstalk with phosphorylation to regulate signaling and transcription in response to nutrients and stress. , 2010, Biochimica et biophysica acta.

[54]  N. Tanaka,et al.  Loss of p53 enhances catalytic activity of IKKβ through O-linked β-N-acetyl glucosamine modification , 2009, Proceedings of the National Academy of Sciences.

[55]  S. Gygi,et al.  Quantitative analysis of complex protein mixtures using isotope-coded affinity tags , 1999, Nature Biotechnology.

[56]  Yup Kang,et al.  Proteomic analysis of O-GlcNAc modifications derived from streptozotocin and glucosamine induced beta-cell apoptosis. , 2007, Journal of biochemistry and molecular biology.

[57]  M. Bardor,et al.  Multiple reaction monitoring mass spectrometry for the discovery and quantification of O-GlcNAc-modified proteins. , 2014, Analytical chemistry.

[58]  Gerald W. Hart,et al.  Glycomics Hits the Big Time , 2010, Cell.

[59]  John R Yates,et al.  Proteomics by mass spectrometry: approaches, advances, and applications. , 2009, Annual review of biomedical engineering.

[60]  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.

[61]  H. Hang,et al.  Chemical reporters for fluorescent detection and identification of O-GlcNAc-modified proteins reveal glycosylation of the ubiquitin ligase NEDD4-1 , 2011, Proceedings of the National Academy of Sciences.

[62]  C. Ha,et al.  Production of O-GlcNAc modified recombinant proteins in Escherichia coli , 2002 .

[63]  Partha Sarathi Banerjee,et al.  Chemoselective attachment of small molecule effector functionality to human adenoviruses facilitates gene delivery to cancer cells. , 2010, Journal of the American Chemical Society.

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

[65]  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.

[66]  G. Hart,et al.  Site‐specific interplay between O‐GlcNAcylation and phosphorylation in cellular regulation , 2010, FEBS letters.

[67]  L. Hsieh‐Wilson,et al.  Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins. , 2008, Journal of the American Chemical Society.

[68]  A. Burlingame,et al.  Polycomb repressive complex 2 is necessary for the normal site-specific O-GlcNAc distribution in mouse embryonic stem cells , 2011, Proceedings of the National Academy of Sciences.

[69]  Nuno Bandeira,et al.  Peptide Identification by Tandem Mass Spectrometry with Alternate Fragmentation Modes* , 2012, Molecular & Cellular Proteomics.

[70]  R. Aebersold,et al.  Selected reaction monitoring for quantitative proteomics: a tutorial , 2008, Molecular systems biology.

[71]  Gerald W. Hart,et al.  O-GlcNAc signalling: implications for cancer cell biology , 2011, Nature Reviews Cancer.

[72]  J. Hanover,et al.  O-GlcNAc cycling: a link between metabolism and chronic disease. , 2013, Annual review of nutrition.

[73]  S. Shibata,et al.  Identification of Ewing's sarcoma gene product as a glycoprotein using a monoclonal antibody that recognizes an immunodeterminant containing O-linked N-acetylglucosamine moiety. , 2002, Hybridoma and hybridomics.

[74]  G. Hart,et al.  O‐GlcNAcylation of Kinases , 2007, Biochemical and biophysical research communications.

[75]  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.

[76]  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.

[77]  P. Kloetzel,et al.  Mapping of O-GlcNAc Sites of 20 S Proteasome Subunits and Hsp90 by a Novel Biotin-Cystamine Tag , 2012, Molecular & Cellular Proteomics.

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

[79]  R. Aebersold,et al.  Simultaneous detection and identification of O-GlcNAc-modified glycoproteins using liquid chromatography-tandem mass spectrometry. , 2000, Analytical chemistry.

[80]  M. Mann,et al.  Stable Isotope Labeling by Amino Acids in Cell Culture, SILAC, as a Simple and Accurate Approach to Expression Proteomics* , 2002, Molecular & Cellular Proteomics.

[81]  L. Heilmeyer,et al.  Determination and location of phosphoserine in proteins and peptides by conversion to S-ethylcysteine. , 1991, Methods in enzymology.

[82]  Jung Weon Lee,et al.  O-GlcNAc modulation at Akt1 Ser473 correlates with apoptosis of murine pancreatic beta cells. , 2008, Experimental cell research.

[83]  K. Medzihradszky,et al.  Enrichment of O-GlcNAc modified proteins by the periodate oxidation-hydrazide resin capture approach. , 2010, Journal of proteome research.

[84]  Bernhard Kuster,et al.  Discovery of O-GlcNAc-modified Proteins in Published Large-scale Proteome Data* , 2012, Molecular & Cellular Proteomics.

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

[86]  Bernhard Kuster,et al.  A Novel Two-Stage Tandem Mass Spectrometry Approach and Scoring Scheme for the Identification of O-GlcNAc Modified Peptides , 2011, Journal of the American Society for Mass Spectrometry.

[87]  Carolyn R. Bertozzi,et al.  Essentials of Glycobiology , 1999 .

[88]  G. Hart,et al.  O-GlcNAc Cycling Enzymes Associate with the Translational Machinery and Modify Core Ribosomal Proteins , 2010, Molecular biology of the cell.

[89]  G. P. Barnard.  A New Ion Source for Mass Spectrometry , 1949, Nature.

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

[91]  W. G. Kelly,et al.  RNA polymerase II is a glycoprotein. Modification of the COOH-terminal domain by O-GlcNAc. , 1993, The Journal of biological chemistry.

[92]  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.

[93]  Brian L. Frey,et al.  Chemical Derivatization of Peptide Carboxyl Groups for Highly Efficient Electron Transfer Dissociation , 2013, Journal of The American Society for Mass Spectrometry.

[94]  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.

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

[96]  A. Burlingame,et al.  Electron transfer dissociation (ETD): The mass spectrometric breakthrough essential for O‐GlcNAc protein site assignments—a study of the O‐GlcNAcylated protein Host Cell Factor C1 , 2013, Proteomics.

[97]  W. G. Kelly,et al.  Glycosylation of chromosomal proteins: Localization of O-linked N-acetylglucosamine in Drosophila chromatin , 1989, Cell.

[98]  J. Michalski,et al.  Identification of substituted sites on MUC5AC mucin motif peptides after enzymatic O-glycosylation combining beta-elimination and fixed-charge derivatization. , 2002, Rapid communications in mass spectrometry : RCM.

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

[100]  M. Mann,et al.  Super-SILAC mix for quantitative proteomics of human tumor tissue , 2010, Nature Methods.

[101]  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.

[102]  M. Buse,et al.  O-Linked N-Acetylglucosamine Modification of Insulin Receptor Substrate-1 Occurs in Close Proximity to Multiple SH2 Domain Binding Motifs* , 2009, Molecular & Cellular Proteomics.

[103]  J. Shabanowitz,et al.  O-GlcNAcylation of the Plum pox virus capsid protein catalyzed by SECRET AGENT: characterization of O-GlcNAc sites by electron transfer dissociation mass spectrometry , 2011, Amino Acids.

[104]  M. Mann,et al.  Quantitative, high-resolution proteomics for data-driven systems biology. , 2011, Annual review of biochemistry.

[105]  A. Gooley,et al.  Identification of glycosylation sites in mucin peptides by edman degradation. , 2000, Methods in molecular biology.

[106]  N. Zachara.  Detecting the "O-GlcNAc-ome"; detection, purification, and analysis of O-GlcNAc modified proteins. , 2009, Methods in molecular biology.

[107]  C. Wright.  Crystal structure of a wheat germ agglutinin/glycophorin-sialoglycopeptide receptor complex. Structural basis for cooperative lectin-cell binding. , 1992, The Journal of biological chemistry.

[108]  Zhi-ping Wang,et al.  O-GlcNAcPRED: a sensitive predictor to capture protein O-GlcNAcylation sites. , 2013, Molecular bioSystems.

[109]  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.

[110]  K. Iqbal,et al.  O-GlcNAc cycling modulates neurodegeneration , 2012, Proceedings of the National Academy of Sciences.

[111]  Gerald W. Hart,et al.  Cycling of O-linked β-N-acetylglucosamine on nucleocytoplasmic proteins , 2007, Nature.

[112]  M. Mann,et al.  Higher-energy C-trap dissociation for peptide modification analysis , 2007, Nature Methods.

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

[114]  A. Burlingame,et al.  Identification of Novel Sites of O-N-Acetylglucosamine Modification of Serum Response Factor Using Quadrupole Time-of-flight Mass Spectrometry* , 2003, Molecular & Cellular Proteomics.

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

[116]  Min-Dian Li,et al.  Cracking the O-GlcNAc code in metabolism , 2013, Trends in Endocrinology & Metabolism.

[117]  K. Parker,et al.  Multiplexed Protein Quantitation in Saccharomyces cerevisiae Using Amine-reactive Isobaric Tagging Reagents*S , 2004, Molecular & Cellular Proteomics.

[118]  R. Neve,et al.  Dynamic O-GlcNAc Modification Regulates CREB-Mediated Gene Expression and Memory Formation , 2011, Nature Chemical Biology.

[119]  G. Hart,et al.  Identification of O-GlcNAc sites on proteins. , 2006, Methods in enzymology.

[120]  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.

[121]  N. Zachara,et al.  Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis , 2013, Cell Stress and Chaperones.

[122]  Ruedi Aebersold,et al.  Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry , 2003, Nature Biotechnology.

[123]  Shu-Hui Chen,et al.  Stable-isotope dimethyl labeling for quantitative proteomics. , 2003, Analytical chemistry.

[124]  Yingming Zhao,et al.  Tagging-via-substrate strategy for probing O-GlcNAc modified proteins. , 2005, Journal of proteome research.

[125]  A. Burlingame,et al.  Identification of GlcNAcylation sites of peptides and alpha-crystallin using Q-TOF mass spectrometry. , 2001, Journal of the American Society for Mass Spectrometry.

[126]  Linda C Hsieh-Wilson,et al.  Phosphofructokinase 1 Glycosylation Regulates Cell Growth and Metabolism , 2012, Science.

[127]  X. Yao,et al.  18O2-labeling in quantitative proteomic strategies: a status report. , 2009, Journal of proteome research.

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

[129]  B. Ramakrishnan,et al.  Structure-based design of beta 1,4-galactosyltransferase I (beta 4Gal-T1) with equally efficient N-acetylgalactosaminyltransferase activity: point mutation broadens beta 4Gal-T1 donor specificity. , 2002, The Journal of biological chemistry.

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

[131]  Hongfang Liu,et al.  dbOGAP - An Integrated Bioinformatics Resource for Protein O-GlcNAcylation , 2011, BMC Bioinformatics.

[132]  L. Gerace,et al.  Monoclonal antibodies identify a group of nuclear pore complex glycoproteins , 1987, The Journal of cell biology.

[133]  G. Hart,et al.  β-N-acetylglucosamine (O-GlcNAc) is part of the histone code , 2010, Proceedings of the National Academy of Sciences.

[134]  M. Bergström,et al.  Bioaffinity chromatography in the 10 mM range of Kd. , 1997, Analytical biochemistry.

[135]  T. Lefebvre,et al.  Identification of Structural and Functional O-Linked N-Acetylglucosamine-bearing Proteins in Xenopus laevis Oocyte*S , 2008, Molecular & Cellular Proteomics.

[136]  G. Hart,et al.  Cross talk between O-GlcNAcylation and phosphorylation: roles in signaling, transcription, and chronic disease. , 2011, Annual review of biochemistry.

[137]  G. Hart,et al.  O‐GlcNAcomics—Revealing roles of O‐GlcNAcylation in disease mechanisms and development of potential diagnostics , 2013, Proteomics. Clinical applications.

[138]  D. Vocadlo,et al.  Mapping O-GlcNAc modification sites on tau and generation of a site-specific O-GlcNAc tau antibody , 2011, Amino Acids.

[139]  M. Mortuaire,et al.  Identification of new O-GlcNAc modified proteins using a click-chemistry-based tagging , 2008, Analytical and bioanalytical chemistry.

[140]  G. Hart,et al.  Nuclear pore complex glycoproteins contain cytoplasmically disposed O- linked N-acetylglucosamine , 1987, The Journal of cell biology.

[141]  W. G. Kelly,et al.  Localization of O-GlcNAc modification on the serum response transcription factor. , 1992, The Journal of biological chemistry.

[142]  G. Hart,et al.  High-sensitivity FAB-MS strategies for O-GlcNAc characterization. , 1991, Glycobiology.

[143]  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.

[144]  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.