Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides.

Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies.

[1]  Jonas Nilsson,et al.  Human Urinary Glycoproteomics; Attachment Site Specific Analysis of N- and O-Linked Glycosylations by CID and ECD* , 2011, Molecular & Cellular Proteomics.

[2]  Targeting the glycoproteome , 2012, Glycoconjugate Journal.

[3]  M F Bean,et al.  Collisional fragmentation of glycopeptides by electrospray ionization LC/MS and LC/MS/MS: methods for selective detection of glycopeptides in protein digests. , 1993, Analytical chemistry.

[4]  Jonathan C Trinidad,et al.  N- and O-Glycosylation in the Murine Synaptosome* , 2013, Molecular & Cellular Proteomics.

[5]  H. Sinn,et al.  Synthetic antitumor vaccines containing MUC1 glycopeptides with two immunodominant domains-induction of a strong immune response against breast tumor tissues. , 2011, Angewandte Chemie.

[6]  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.

[7]  C. Sihlbom,et al.  Identification of Chondroitin Sulfate Linkage Region Glycopeptides Reveals Prohormones as a Novel Class of Proteoglycans* , 2014, Molecular & Cellular Proteomics.

[8]  Michael Przybylski,et al.  Elucidation of O-glycosylation structures of the beta-amyloid precursor protein by liquid chromatography-mass spectrometry using electron transfer dissociation and collision induced dissociation. , 2009, Journal of proteome research.

[9]  Martin R Larsen,et al.  Selective enrichment of sialic acid–containing glycopeptides using titanium dioxide chromatography with analysis by HILIC and mass spectrometry , 2010, Nature Protocols.

[10]  E. Schmitt,et al.  Synthetic vaccines consisting of tumor-associated MUC1 glycopeptide antigens and a T-cell epitope for the induction of a highly specific humoral immune response. , 2008, Angewandte Chemie.

[11]  Liping Yu,et al.  O-Mannosyl Phosphorylation of Alpha-Dystroglycan Is Required for Laminin Binding , 2010, Science.

[12]  Koichi Tanaka,et al.  Fragmentation Characteristics of Deprotonated N-linked Glycopeptides: Influences of Amino Acid Composition and Sequence , 2014, Journal of The American Society for Mass Spectrometry.

[13]  R. Cummings,et al.  The Tn Antigen—Structural Simplicity and Biological Complexity , 2011, Angewandte Chemie.

[14]  J. Leary,et al.  Preparation, separation, and conformational analysis of differentially sulfated heparin octasaccharide isomers using ion mobility mass spectrometry. , 2012, Analytical chemistry.

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

[16]  Christian Pett,et al.  A convergent strategy for the synthesis of type-1 elongated mucin cores 1-3 and the corresponding glycopeptides. , 2014, Chemistry.

[17]  C. Hesse,et al.  Enrichment of glycopeptides for glycan structure and attachment site identification , 2009, Nature Methods.

[18]  G. Larson,et al.  LC-MS/MS characterization of O-glycosylation sites and glycan structures of human cerebrospinal fluid glycoproteins. , 2013, Journal of proteome research.

[19]  Yoshiki Narimatsu,et al.  The GalNAc-type O-Glycoproteome of CHO Cells Characterized by the SimpleCell Strategy* , 2014, Molecular & Cellular Proteomics.

[20]  A. Suzuki,et al.  Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry. , 2008, Analytical chemistry.

[21]  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.

[22]  H. Wandall,et al.  Mining the O-glycoproteome using zinc-finger nuclease–glycoengineered SimpleCell lines , 2011, Nature Methods.

[23]  K. Furukawa,et al.  O-Linked N-Acetylglucosamine Is Present on the Extracellular Domain of Notch Receptors* , 2008, Journal of Biological Chemistry.

[24]  A. Westman-Brinkmalm,et al.  Tandem Mass Spectrometry , 2008 .

[25]  Toshinori Sato,et al.  The distinction of underivatized monosaccharides using electrospray ionization ion trap mass spectrometry. , 2007, Rapid communications in mass spectrometry : RCM.

[26]  R. Zubarev,et al.  Localization of O-glycosylation sites in peptides by electron capture dissociation in a Fourier transform mass spectrometer. , 1999, Analytical chemistry.

[27]  H. Cooper,et al.  Separation and Identification of Isomeric Glycopeptides by High Field Asymmetric Waveform Ion Mobility Spectrometry , 2012, Analytical chemistry.

[28]  She Chen,et al.  Pathogen blocks host death receptor signalling by arginine GlcNAcylation of death domains , 2013, Nature.

[29]  H. Waldmann,et al.  Tumor-associated MUC1 tandem-repeat glycopeptide microarrays to evaluate serum- and monoclonal-antibody specificity. , 2009, Angewandte Chemie.

[30]  K. Medzihradszky,et al.  How to Dig Deeper? Improved Enrichment Methods for Mucin Core-1 Type Glycopeptides* , 2012, Molecular & Cellular Proteomics.

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

[32]  Suh-Yuen Liang,et al.  Sweet-Heart - an integrated suite of enabling computational tools for automated MS2/MS3 sequencing and identification of glycopeptides. , 2013, Journal of proteomics.

[33]  Daniel S Spellman,et al.  Quantitative analysis of intact apolipoproteins in human HDL by top-down differential mass spectrometry , 2010, Proceedings of the National Academy of Sciences.

[34]  C. Sihlbom,et al.  Evaluation of the combination of bead technology with SELDI-TOF-MS and 2-D DIGE for detection of plasma proteins. , 2008, Journal of proteome research.

[35]  E. Bennett,et al.  Mining the O-mannose glycoproteome reveals cadherins as major O-mannosylated glycoproteins , 2013, Proceedings of the National Academy of Sciences.

[36]  Nicolle H Packer,et al.  Advances in LC-MS/MS-based glycoproteomics: getting closer to system-wide site-specific mapping of the N- and O-glycoproteome. , 2014, Biochimica et biophysica acta.

[37]  André M Deelder,et al.  Cotton HILIC SPE microtips for microscale purification and enrichment of glycans and glycopeptides. , 2011, Analytical chemistry.

[38]  M. Raftery,et al.  Glycosylation and Sialylation of Macrophage-derived Human Apolipoprotein E Analyzed by SDS-PAGE and Mass Spectrometry , 2010, Molecular & Cellular Proteomics.

[39]  Martin R Larsen,et al.  Titanium dioxide as chemo-affinity chromatographic sorbent of biomolecular compounds--applications in acidic modification-specific proteomics. , 2011, Journal of proteomics.

[40]  K. Blennow,et al.  Site-specific characterization of threonine, serine, and tyrosine glycosylations of amyloid precursor protein/amyloid β-peptides in human cerebrospinal fluid , 2011, Proceedings of the National Academy of Sciences.

[41]  Manfred Wuhrer,et al.  Glycomics using mass spectrometry , 2012, Glycoconjugate Journal.

[42]  Nichollas E. Scott,et al.  Site-specific glycan-peptide analysis for determination of N-glycoproteome heterogeneity. , 2013, Journal of proteome research.

[43]  D. Harvey.  Derivatization of carbohydrates for analysis by chromatography; electrophoresis and mass spectrometry. , 2011, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[44]  A. Beck,et al.  Middle-down analysis of monoclonal antibodies with electron transfer dissociation orbitrap fourier transform mass spectrometry. , 2014, Analytical chemistry.

[45]  G. Larson,et al.  Sialic acid capture-and-release and LC-MS(n) analysis of glycopeptides. , 2013, Methods in molecular biology.

[46]  E. Suzuki,et al.  O-linked-N-acetylglucosamine on extracellular protein domains mediates epithelial cell-matrix interactions. , 2011, Nature communications.

[47]  Christian Pett,et al.  A unified strategy for the synthesis of mucin cores 1-4 saccharides and the assembled multivalent glycopeptides. , 2013, Chemistry.

[48]  Jonas Nilsson,et al.  Characterization of site-specific O-glycan structures within the mucin-like domain of alpha-dystroglycan from human skeletal muscle. , 2010, Glycobiology.

[49]  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.

[50]  K. Medzihradszky.  Characterization of protein N-glycosylation. , 2005, Methods in enzymology.

[51]  D. Ashline,et al.  Congruent strategies for carbohydrate sequencing. 1. Mining structural details by MSn. , 2005, Analytical chemistry.

[52]  S. Brunak,et al.  Precision mapping of the human O‐GalNAc glycoproteome through SimpleCell technology , 2013, The EMBO journal.

[53]  G. Larson,et al.  O-Mannose and O-N-acetyl galactosamine glycosylation of mammalian α-dystroglycan is conserved in a region-specific manner. , 2012, Glycobiology.

[54]  M. Rejžek,et al.  Discrimination of epimeric glycans and glycopeptides using IM-MS and its potential for carbohydrate sequencing , 2013, Nature Chemistry.

[55]  J. Peter-Katalinic,et al.  Methods in enzymology: O-glycosylation of proteins. , 2005, Methods in enzymology.