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The majority of all proteins undergo posttranslational modifications that significantly alter their physical and chemical properties, including their folding and conformation distribution, their stability, and, consequently, their activity and function. In Posttranslational Modifications of Proteins: Tools for Functional Proteomics, Christoph Kannicht and a panel of highly experienced researchers describe readily reproducible methods for detecting and analyzing the most important of these modifications, particularly with regard to protein function, proteome research, and the characterization of pharmaceutical proteins. Among the methods presented are those for analyzing the assignment of disulfide bond sites in proteins, protein N-glycosylation and protein O-glycosylation, and oligosaccharides present at specific single glycosylation sites in a protein. Additional powerful techniques facilitate the analysis of glycosylphosphatidylinositols, lipid modifications, protein phosphorylation and sulfation, protein methylation and acetylation, a-amidation, g-glutamate, isoaspartate, and lysine hydroxylation.
Comprehensive and state-of-the-art, Posttranslational Modifications of Proteins: Tools for Functional Proteomics serves as a highly practical guide for all investigators of protein structure-function relationships not only in chemical and pharmaceutical research, but also throughout the rapidly growing field of functional proteomics.
List of contents
Assignment of Disulfide Bonds in Proteins by Chemical Cleavage and Peptide Mapping by Mass Spectrometry.- Carbohydrate Composition Analysis of Glycoproteins Using Highly Sensitive Fluorescence Detection Methods.- Enzymatical Hydrolysis of N-Glycans from Glycoproteins and Fluorescent Labeling by 2-Aminobenzamide (2-AB).- Separation of N-Glycans by HPLC.- Enzymatic Sequence Analysis of N-Glycans.- Immunological Detection of O-GlcNAc.- Analysis of O-Glycosylation.- Characterization of Site-Specific Glycosylation.- Monitoring Glycosylation of Therapeutic Glycoproteins for Consistency Using Highly Fluorescent Anthranilic Acid.- Metabolic Labeling and Structural Analysis of Glycosylphosphatidylinositols from Parasitic Protozoa.- Analysis of S-Acylation of Proteins.- Immunoblotting Methods for the Study of Protein Ubiquitination.- Analysis of Methylation and Acetylation in E. coli Ribosomal Proteins.- Identification of In Vivo Protein Phosphorylation Sites with Mass Spectrometry.- Analysis of Tyrosine-O-Sulfation.- ?-Amidated Peptides: Approaches for Analysis.- ?-Glutamate and p-Hydroxyaspartate in Proteins.- Detection of isoAspartate Residues as a Posttranslational Modification of Proteins and Peptides.- Lysine Hydroxylation and Crosslinking of Collagen.- Heterologous Expression in Endocrine Cells for Analysis of Posttranslational Modifications.- 2D-Electrophoresis: Detection of Glycosylation and Influence on Spot Pattern.
Summary
Describes reproducible methods for detecting and analyzing the posttranslational modifications of protein, particularly with regard to protein function, proteome research, and the characterization of pharmaceutical proteins. This work also presents methods for analyzing the assignment of disulfide bond sites in proteins.
Additional text
"As indicated by the editor, this book will be useful to researchers involved in elucidating protein structure and function as well as those working to design proteins with specific function...this book offers detailed analytical protocols for identifying and characterizing most types of co- and post- translational modifications" - Pharmaceutical Research
"This book provides researchers in the biological sciences with the most updated methods to investigate co- and/or post-translational modifications of proteins...Either trained professionals and students working in the field of proteomics can take benefit for their research." - Proteomics
"It will definitely find appreciative readers among a wide audience of glycobiologists, biochemists, molecular biologists, biotechnologists, and researchers specializing in pharmacology and molecular medicine. This book will be a very useful tool for teachers and students of universities and institutes." -Biochemistry
Report
"As indicated by the editor, this book will be useful to researchers involved in elucidating protein structure and function as well as those working to design proteins with specific function...this book offers detailed analytical protocols for identifying and characterizing most types of co- and post- translational modifications" - Pharmaceutical Research
"This book provides researchers in the biological sciences with the most updated methods to investigate co- and/or post-translational modifications of proteins...Either trained professionals and students working in the field of proteomics can take benefit for their research." - Proteomics
"It will definitely find appreciative readers among a wide audience of glycobiologists, biochemists, molecular biologists, biotechnologists, and researchers specializing in pharmacology and molecular medicine. This book will be a very useful tool for teachers and students of universities and institutes." -Biochemistry
