Posttranslational Modifications of Proteins: Tools for Functional Proteomics (Methods in Molecular Biology Vol 194)

✍ Scribed by Christoph Kannicht (Ed.)

• Year: 2002
• Tongue: English
• Leaves: 336
• Edition: 1st
• Category: Library

✦ Synopsis

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.

✦ Table of Contents

Front......Page 1
Preface......Page 6
Contents......Page 8
Contributors......Page 10
1 Assignment of Disulfi de Bonds in Proteins by Chemical Cleavage and Peptide Mapping by Mass Spectrometry......Page 14
2 Carbohydrate Composition Analysis of Glycoproteins Using Highly Sensitive Fluorescence Detection Methods......Page 36
3 Enzymatical Hydrolysis of N-Glycans from Glycoproteins and Fluorescent Labeling by 2-Aminobenzamide (2-AB)......Page 50
4 Separation of N-Glycans by HPLC......Page 58
5 Enzymatic Sequence Analysis of N-Glycans......Page 76
6 Immunological Detection of O-GlcNAc......Page 86
7 Analysis of O-Glycosylation......Page 102
8 Characterization of Site-Specifi c Glycosylation......Page 114
9 Monitoring Glycosylation of Therapeutic Glycoproteins for Consistency Using Highly Fluorescent Anthranilic Acid......Page 140
10 Metabolic Labeling and Structural Analysis of Glycosylphosphatidylinositols from Parasitic Protozoa......Page 156
11 Analysis of S-Acylation of Proteins......Page 172
12 Immunoblotting Methods for the Study of Protein Ubiquitination......Page 192
13 Analysis of Methylation and Acetylation in E. coli Ribosomal Proteins......Page 218
14 Identifi cation of In Vivo Protein Phosphorylation Sites with Mass Spectrometry......Page 224
15 Analysis of Tyrosine-O-Sulfation......Page 236
16 α-Amidated Peptides......Page 254
17 γ-Glutamate and β-Hydroxyaspartate in Proteins......Page 272
18 Detection of isoAspartate Residues as a Posttranslational Modifi cation of Proteins and Peptides......Page 282
19 Lysine Hydroxylation and Crosslinking of Collagen......Page 290
20 Heterologous Expression in Endocrine Cells for Analysis of Posttranslational Modifications......Page 304
21 2D-Electrophoresis......Page 314
Index......Page 330
Back......Page 336