"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.

I Separation of Proteins

1 Purification of Proteins for Sequencing
2 Affinity Chromatography of Proteins
3 Protein Purification by Aqueous Two-Phase Systems
4 Rapid HPLC Separation of Proteins, Peptides, and Amino Acids Using Short Columns Filled with Nonporous Silica-Based Particles
II Separation of Peptides
5 Enzymatic and Chemical Cleavages of Proteins
6 Separation of Peptides Using HPLC and TLC Fingerprints
7 Microseparation of In Situ Digested Peptides
8 Isolation of Peptides for Microsequencing by In-Gel Proteolytic Digestion
III Manual and Automated Microsequencing Methods
9 Automated Microsequencing: Introduction and Overview
10 A Manual Method for Protein Sequence Analysis Using the DABITC/PITC Method
11 Solid-Phase Sequencing of Peptides and Proteins
12 Sequence Analysis of the NH2-Terminally Blocked Proteins Immobilized on PVDF Membranes from Polyacrylamide Gels
IV Electrophoresis and Blotting of Proteins
13 Two-Dimensional Electrophoresis
14 Semi-Dry Blotting onto Hydrophobic Membranes
V Analysis of Amino Acids
15 Highly Sensitive Amino Acid Analysis
16 Quantitative Analysis of D- and L-Amino Acids by HPLC
VI Identification of Cysteine Residues and Lipids in Proteins
17 Identification of Cysteine Residues and Disulfide Bonds in Proteins
18 Lipid Modifications of Proteins
VII Mass Spectrometry in Peptide and Protein Analysis
19 Mass Spectrometry in Peptide and Protein Sequence Analysis
20 Carboxy-Terminal Sequencing by Combining Carboxypeptidase Y and P Digestion with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
21 Crystallization of Biological Macromolecules.