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Glycosylation is the most abundant post-translational modification of proteins. Estimates vary widely, but a common assessment is that upwards of 50 per cent of eukaryotic proteins are modified by some type of glycan. In Mass Spectrometry of Glycoproteins: Methods and Protocols, expert researchers in the field detail many of the methods that are now commonly used for glycoproteomics. These methods and techniques include robust sample preparation techniques; advanced chromatographic strategies for improving dynamic range; state-of-the-art mass spectrometry instrumentation and associated ionization and fragmentation methods; and informatics tools used for identifying glycoproteins and characterizing the associated glycans.
Written in the highly successful Methods in Molecular Biology format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Mass Spectrometry of Glycoproteins: Methods and Protocols is an essential resource for those who work at the interface of glycobiology and mass spectrometry.
1. Introduction to Glycosylation and Mass Spectrometry Steven M. Patrie, Michael J. Roth and Jennifer J. Kohler P
Part I - Enrichment and Isolation Methods
2. Tandem Lectin Weak Affinity Chromatography (LWAC) for Glycoprotein Enrichment Zhi Yuan Ma, Yuliya Skorobogatko, and Keith Vosseller
3. CSC Technology - Selective Labeling of Glycoproteins by Mild Oxidation to Phenotype Cells Andreas Hofmann,Damaris Bausch-Fluck, and Bernd Wollscheid
4. Use of Boronic Acid Nano
particles in Glycoprotein Enrichment Yawei Xu, Lijuan Zhang and Haojie Lu
5. Incorporation of Unnatural Sugars for the Identification of Glycoproteins Balyn W. Zaro , Howard C. Hang , Matthew R. Pratt
6. Characterization of Membrane-associated Glycoproteins using Lectin Affinity Chromatography and Mass Spectrometry Yashu Liu, Jintang He and David M. Lubman
7. Sialic Acid Capture-and-Release and LC-MSn Analysis of Glycopeptides Jonas Nilsson and Goran Larson
Part II - Sample Preparation
8. In-Solution Digestion of Glycoproteins for Glycopeptide-Based Mass Analysis Eden P. Go, Kathryn R. Rebecchi, and Heather Desaire
9. Nano-HPLC-MS of Glycopeptides Obtained After Non-specific Proteolysis Gerhild Zauner, Carolien A. M. Koeleman, Andre M. Deelder and Manfred Wuhrer
Part III - Separation Methods
10. Glycopeptide Enrichment for MALDI-TOF Mass Spectrometry Analysis by Hydrophilic Interaction Liquid Chromatography Solid Phase Extraction (HILIC SPE) Pia Honnerup Jensen, Simon Mysling, Peter Hojrup and Ole Norregaard Jensen
11. Separation and Identification of Glycoforms by Capillary Electrophoresis with Electrospray Ionization Mass Spectrometric Detection Alina D. Zamfir , Corina Flangea, Alina Serb3, Ana-Maria Zagrean4, Andreas M. Rizzi5 and Eugen Sisu
12. Structural Separations by Ion Mobility-MS for Glycomics and Glycoproteomics Larissa S. Fenn and John A. McLean
Part IV - Quantitation Methods
13. Quantitative Analysis of Glycoprotein Glycans Ron Orlando
14. Stable Isotope Labeling of N-glycosylated Peptides by Enzymatic Deglycosylation for Mass Spectrometry-based Glycoproteomics Hiroyuki Kaji and Toshiaki Isobe
15. Approaches for Site Mapping and Quantification of O-linked Glycopeptides Peng Zhao, Stephanie H. Stalnaker, Lance Wells
16. Glycan Profiling: Label-free Analysis of Glycoproteins Yoshinao Wada
Part V - Computational Tools
17. Introduction to Informatics in Glycoprotein Analysis Kiyoko F. Aoki-Kinoshita
18. Software Tools for Glycan Profiling Chuan-Yih Yu, Anoop Mayampurath, Haixu Tang
Part VI - Case Studies in Mass Spectrometry of Glycoproteins
19. Quantitative Characterization of Glycoproteins in Neurodegenerative Disorders using iTRAQ Min Shi, Hyejin Hwang and Jing Zhang
20. Quantitative Proteomic Analysis of N-linked Glycoproteins in Human Tear FluidLei Zhou and Roger W. Beuerman
21. Elucidation of N-glycosites Within Human Plasma Glycoproteins for Cancer Biomarker Discovery Penelope Drake, Birgit Schilling, Brad Gibson and Susan Fisher
22. Characterizing the Glycosylation State of Therapeutic Recombinant Glycoproteins Nicole Samuels, David Kates, Jun Liu, Joanne Severs
