Protein Sequencing Protocols: (64 Methods in Molecular Biology)

A timely collection of current and state-of-the-art techniques for protein and peptide preparation and sequencing - all described by hands-on, masters of the procedures. Conveys valuable lessons and insights into using automated sequencers and mass spectrometers, preparing proteins and peptides on a microgram and submicrogram scale, mapping peptides, analyzing amino acids by pre- and post-column derivatization methods, and sequencing N- and C-terminal peptides by classical and newly developed techniques. Also deals with such troublesome problems as identifying sites of modification and working with very small samples.

Table of Contents:
Strategies for Handling Polypeptides on a Microscale Bryan John Smith and Paul Tempst. SDS Polyacrylamide Gel Electrophoresis for N-Terminal Protein Sequencing Bryan John Smith. Two-Dimensional Polyacrylamide Gel Electrophoresis for the Separation of Proteins for Chemical Characterization Michael J. Dunn. Electroblotting of Proteins from Polyacrylamide Gels for Chemical Characterization Michael J. Dunn. Enzymatic Cleavage of Polypeptides Bryan John Smith and Colin Wheeler. Chemical Cleavage of Polypeptides Bryan John Smith. Preparation of Glycopeptides Chris W. Sutton and Jacqui A. O'Neill. Selective Isolation of the Carboxy Terminal Peptide from a Protein Tomoko Hayashi Akiyama and Tatsuru Sasagawa. Selective Isolation of the Amino Terminal Peptide from a-Amino Blocked Protein Tomoko Hayashi Akiyama and Tatsuru Sasagawa. Analytical and Micropreparative Capillary Electrophoresis of Peptides Alan J. Smith. Reverse-Phase HPLC Purification of Peptides from Natural Sources for Structural Analysis Chris Shaw . High-Performance Liquid Chromatography On-Line Derivative Spectroscopy for the Characterization of Peptides with Aromatic Amino Acid Residues Christoph W. Turck. Hydrolysis of Samples for Amino Acid Analysis Ian Davidson. Amino Acid Analysis: Precolumn Derivatization Methods G. Brent Irvine. Postcolumn Amino Acid Analysis Alan J. Smith. Fast-Atom Bombardment and Electrospray Mass Spectrometry of Peptides Proteins and Glycoproteins Fiona M. Greer and Howard R. Morris . Peptide Mass Fingerprinting Using MALDI-TOF Mass Spectrometry Darryl J. C. Pappin. N-Terminal Protein Ladder Sequencing Rong Wang and Brian T. Chait. The Dansyl-Edman Method for Manual Peptide Sequencing John M. Walker. The Dansyl Method for Identifying N-Terminal Amino Acids John M. Walker. Identification of PTH-Amino Acids by High-Performance Liquid Chromatography Gregory A. Grant and Mark W. Crankshaw. Protein Sequencer Maintenance and Troubleshooting Bryan Dunbar. Identifying the Carboxy Terminal Amino Acid of a Protein Tomoko Hayashi Akiyama and Tatsuru Sasagawa. Enzymatic and Chemical Methods for Manual C-Terminal Peptide Sequencing Franca Casagranda and John F. K. Wilshire. Automated Methods for C-Terminal Protein Sequencing Jerome M. Bailey and Chad G. Miller. Chemical Modification of Proteins for Sequence Analysis Alex F. Carne and Sally U. On-Membrane Deblocking of Proteins Hisashi Hirano Setsuko Komatsu and Susumu Tsunasawa. Analysis of Sites of Protein Phosphorylation Alastair Aitken and MichSle Learmonth. Identification of Sites of Glycosylation Chris W. Sutton and Jacqui A. O'Neill. Quantification and Location of Disulfide Bonds in Proteins Alastair Aitken and MichSle Learmonth. Appendix 1: Letter Codes Structures Masses and Derivatives of Amino Acids James P. Turner and Bryan John Smith. Appendix 2: A Library of Consensus Sequences Alastair Aitken. Index.

Review :
This methodological guide can be recommended to all who are concerned with protein characterization or peptide mapping. - Reviewed by R. Moravec, Biologia Plantarum