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Kannicht (Institut fur Molekularbiologie und Biochemie, Freie Universitat Berlin, Germany) presents a compilation of protocols needed to detect and analyze important co- and posttranslational modification of proteins. Focusing on the fields of protein function, proteome research, and the characterization of pharmaceutical proteins, chapters provide step-by-step instructions on the analysis of such protein modifications as the assignment of disulfide bond sites in proteins, protein glycosylation, carbohydrate composition, the cleavage and separation of N-linked glycans, and detection of O-linked N-acetylglucosamine residues.Book News, Inc.

Book Info

Freie Universitat Berlin, Germany. Features reproducible methods for detecting and analyzing posttranslational modifications of proteins, instructions ensuring results, and tips on avoiding pitfalls. Emphasizes protein function, proteome research, and the characterization of pharmaceutical proteins.

Book Description

Christoph Kannicht and a panel of highly experienced researchers describe readily 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. 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.

Preface
Contributors
1 Assignment of Disulfide Bonds in Proteins by Chemical Cleavage and Peptide Mapping by Mass Spectrometry 1
2 Carbohydrate Composition Analysis of Glycoproteins Using Highly Sensitive Fluorescence Detection Methods 23
3 Enzymatical Hydrolysis of N-Glycans from Glycoproteins and Fluorescent Labeling by 2-Aminobenzamide (2-AB) 37
4 Separation of N-Glycans by HPLC 45
5 Enzymatic Sequence Analysis of N-Glycans 63
6 Immunological Detection of O-GlcNAc 73
7 Analysis of O-Glycosylation 89
8 Characterization of Site-Specific Glycosylation 101
9 Monitoring Glycosylation of Therapeutic Glycoproteins for Consistency Using Highly Fluorescent Anthranilic Acid 127
10 Metabolic Labeling and Structural Analysis of Glycosylphosphatidylinositols from Parasitic Protozoa 143
11 Analysis of S-Acylation of Proteins 159
12 Immunoblotting Methods for the Study of Protein Ubiquitination 179
13 Analysis of Methylation and Acetylation in E. coli Ribosomal Proteins 205
14 Identification of In Vivo Protein Phosphorylation Sites with Mass Spectrometry 211
15 Analysis of Tyrosine-O-Sulfation 223
16 [alpha]-Amidated Peptides: Approaches for Analysis 241
17 [gamma]-Glutamate and [beta]-Hydroxyaspartate in Proteins 259
18 Detection of isoAspartate Residues as a Posttranslational Modification of Proteins and Peptides 269
19 Lysine Hydroxylation and Crosslinking of Collagen 277
20 Heterologous Expression in Endocrine Cells for Analysis of Posttranslational Modifications 291
21 2D-Electrophoresis: Detection of Glycosylation and Influence on Spot Pattern 301
Index 317