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The majority of all proteins undergo co- and/or post-translational modifications, crucially altering physical and chemical properties, folding, conformation distribution, stability, activity, and, consequently, the function of the proteins. In Post-translational Modifications of Proteins: Tools for Functional Proteomics, Second Edition, this important area of research is brought up-to-date by the leading scientists in the field. This compilation of detailed protocols focuses on protein function, proteome research and characterization of pharmaceutical proteins, while following the successful format of the Methods in Molecular Biology TM series. Each chapter provides a brief introduction to the topic, step-by-step laboratory protocols, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting edge, Post-translational Modifications of Proteins: Tools for Functional Proteomics, Second Edition serves as practical guide for researchers working in the field of protein structure-function relationships and the rapidly growing field of proteomics, as well as scientists in the pharmaceutical industries.
Contents Preface Contributors 1 Disulfide Bond Mapping by Cyanylation-Induced Cleavage and Mass Spectrometry Jiang Wu 2 Detection of Post-Translational Modifications by Fluorescent Staining of 2D gels Archana M. Jacob and Chris W. Turck 3 Identification of Protein Phosphorylation Sites by Advanced LC-ESI-MS/MS Methods Christoph Weise and Christof Lenz 4 Analysis of Tyrosine-O-Sulfation Jens R. Bundgaard, Jette W. Sen, Anders H. Johnsen, and Jens F. Rehfeld 5 a -Amidated Peptides: Approaches for Analysis Gregory P. Mueller and William J. Driscoll 6 Gamma-Glutamate and b -Hydroxyaspartate in Proteins Francis J. Castellino, Victoria A. Ploplis, and Li Zhang 7 Lysine Hydroxylation and Cross-linking of Collagen Mitsuo Yamauchi and Masashi Shiiba 8 Mass Spectrometric Determination of Protein Ubiquitination Carol E. Parker, Maria R. E. Warren, Viorel Mocanu, Susanna F. Greer, and Christoph H. Borchers 9 Analysis of Sumoylation Andrea Pichler 10 Detection and Analysis of Protein ISGylation Tomoharu Takeuchi and Hideyoshi Yokosawa 11 Analysis of Methylation, Acetylation and other Modifications in Bacterial Ribosomal Proteins Randy J. Arnold, William Running, and James P. Reilly 12 Analysis of S-Acylation of Proteins Michael Veit, Evgeni Ponimaskin, and Michael F.G. Schmidt 13 Metabolic Labeling and Structural Analysis of Glycosylphosphatidylinositols from Parasitic Protozoa Nahid Azzouz, Peter Gerold, and Ralph T. Schwarz 14 2D-Electrophoresis: Detection of Glycosylation and Influence on Spot Pattern Klemens Loster and Christoph Kannicht 15 Carbohydrate Composition Analysis of Glycoproteins by HPLC Using Highly Fluorescent Anthranilic Acid (AA) Tag George N. Saddic, Shirish T. Dhume, and Kalyan R. Anumula 16 Enzymatical Hydrolysis of N-glycans from Glycoproteins and Fluorescent Labelling by 2-Aminobenzamide (2-AB) Rolf Nuck 17 Separation of N-glycans by HPLC Martin Gohlke and Veronique Blanchard 18 Enzymatic Sequence Analysis of N-glycans By Exoglycosidase Cleavage and Mass Spectrometry -- Detection of Lewis X Structures Christoph Kannicht, Detlef Grunow, and Lothar Lucka 19 Immunochamical Methods for the Rapid Sceening of the O-Glycosidically Linked N-Acetylglucosamine Modification of Proteins Monika Rex-Mathes, Jurgen Koch, Sabine Werner, Lee S. Griffith, and Brigitte Schmitz 20 Analysis of O-Glycosylation Juan J. Calvete and Libia Sanz 21 Characterization of Site Specific N-Glycosylation Katalin F. Medzihradszky 22 Monitoring Glycosylation of Therapeutic Glycoproteins for Consistency by HPLC Using Highly Fluorescent Anthranilic Acid (AA) Tag Shirish T. Dhume, George N. Saddic, and Kalyan R. Anumula 23 Comparability and Monitoring Immunogenic N-linked Oligosaccharides from Recombinant Monoclonal Antibodies from two Different Cell Lines using HPLC with Fluorescence Detection and Mass Spectrometry Bruce R. Kilgore, Adam W. Lucka, Rekha Patel, Bruce A. Andrien, Jr., and Shirish T. Dhume 24 Mass Spectrometry and HPLC with Fluorescent Detection-Based Orthogonal Approaches to Characterize N-linked Oligosaccharides of Recombinant Monoclonal Antibodies Adam Lucka, Bruce R. Kilgore, Rekha Patel, Bruce A. Andrien, Jr., and Shirish T. Dhume 25 Web-based Computational Tools for the Prediction and Analysis of Posttranslational Modifications of Proteins Vladimir A. Ivanisenko, Dimitry A. Afinnikov, and Nikolay A. Kolchanov