Protein Sequencing Protocols

ISBN-13: 9781617372841 / Angielski / Miękka / 2010 / 493 str.

Bryan John Smith

Opis

Determination of the protein sequence is as important today as it was a half century ago, even though the techniques and purposes have changed over time. Mass spectrometry has continued its recent rapid development to find notable application in the characterization of small amounts of protein, for example, in the field of proteomics. The traditional chemical N-terminal sequencing is still of great value in quality assurance of the increasing number of biopharmaceuticals that are to be found in the clinic, checking processing events of recombinant proteins, and so on. It is joined in the armory of me- ods of protein analysis by such techniques as C-terminal sequencing and amino acid analysis. These methods are continually developing. The first edition of Protein Sequencing Protocols was a snapshot of methods in use in protein biochemistry laboratories at the time, and this, the second edition, is likewise. Methods have evolved in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches. Thus, in this edition, there is inclusion of approaches to validation of methods for quality assurance work, reflecting the current importance of biopharmaceuticals, and also a guide to further analysis of protein sequence information, acknowledging the importance of bioinformatics."

Recenzje

"...this book would certainly benefit readers who are new to the field as well as those more experienced as it serves as a convenient collection of tried and trusted methods...the book gives valuable background and insight to the various techniques associated with protein sequencing, which are often used by those providing services from within a core facility to the wider population of life science researchers." -Proteomics

"A positive feature of this book is that the fact that minute details, which are essential for successful implementation of an analytical method are provided. This book can serve as a useful reference for laboratories which mainly utilize chemical methods for protein analysis." -Pharmaceutical Research

From Reviews of the First Edition...

"...full of valuable information..." -Biochemical Education
"...provides very useful resources for those embarking on protein identification or characterization projects..." -FEBS Letters
"...an extremely useful compendium of protein analysis protocols..." -Nahrung-Food
"...well organized and accessible..." -Journal of the American Chemical Society

"The overall result is a concise and highly informative laboratory manual aimed at both operators and users of protein analysis facilities providing an understanding of what techniques involve, how samples should be prepared and in what form and amounts they should be submitted to achieve the most successful data. This publication has certainly proved to be a valuable asset to my laboratory and I would recommend it not only to those in protein sequencing and mass spectrometry laboratories, but also anyone with protein chemistry applications and interests."-Molecular Biotechnology

Spis treści

Strategies for Handling Polypeptides on a Micro-Scale 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 Proteins Bryan John Smith Chemical Cleavage of Polypeptides Bryan John Smith Enzymatic Preparation and Isolation of Glycopeptides David J. Harvey Analytical and Micropreparative Capillary Electrophoresis of Peptides Alan J. Smith 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 and Ian Davidson Post Column Amino Acid Analysis Alan J. Smith Amino Acid Analysis Using Pre-Column Derivatization with 6-Aminoquinolyl-N-Hydroxysuccinimidyl Carbamate: Analysis of Hydrolyzed Proteins and Electroblotted Samples Steven A. Cohen Amino Acid Analysis in Protein Hydrolysates Using Anion Exchange Chromatography and IPAD Detection Petr Jandik, Jun Cheng, and Nebojsa Avdalovic d-Amino Acid Analysis Andrea Scaloni, Maurizio Simmaco, and Francesco Bossa Validation of Amino Acid Analysis Methods Andrew J. Reason 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 Nanoelectrospray Tandem Mass Spectrometry and Sequence Similarity Searching for Identification of Proteins from Organisms with Unknown Genomes Anna Shevchenko, Shamil Sunyaev, Adam Liska, Peer Bork, and Andrej Shevchenko Direct Identification of Proteins in Ultracomplex Mixtures: Applications to Proteome Analysis David M. Schieltz and John R. Yates, III Identification of PTH-Amino Acids by HPLC Gregory A. Grant and Mark W. Crankshaw Protein Sequencer Maintenance and Troubleshooting Bryan Dunbar N-Terminal Protein Sequencing for Special Applications Philip J. Jackson Identification of Phosphorylation Sites by Edman Degradation Laurey Steinke and Richard G. Cook Validation of Protein Sequencing in a Regulated Laboratory Philip J. Jackson and Neil Dodsworth Automated C-Terminal Protein Sequence Analysis Using the Alkylated-Thiohydantoin Method David R. Dupont, Sylvia W. Yuen, and Kenneth S. Graham Chemical Modifications of Proteins as an Aid to Sequence Analysis Alex F. Carne Deblocking of N-Terminally Modified Proteins Hisashi Hirano and Roza M. Kamp Deblocking of Proteins Containing N-Terminal Pyroglutamic Acid Jacek Mozdzanowski Identification of Sites of Protein Glycosylation David J. Harvey Analysis of Sites of Protein Phosphorylation Alastair Aitken and Michele Learmonth Quantitation and Location of Disulfide Bonds in Proteins Alastair Aitken and Michele Learmonth Getting the Most from Your Protein Sequence Richard R. Copley and Robert B. Russell Appendix 1: Letter Codes, Structures, Masses, and Derivatives of Amino Acids James P. Turner Appendix 2: Protein Consensus Sequence Motifs Alastair Aitken Index

Blurb

Bryan John Smith updates his much-acclaimed first edition of Protein Sequencing Protocols with new and refreshed techniques for determining the sequence of proteins and peptides. This edition includes not only novel approaches to the validation of quality assurance methods, reflecting the current importance of biopharmaceuticals, but also offers a guide to the analysis of protein sequence information via the powerful new tools of bioinformatics. Each readily reproducible protocol contains detailed step-by-step instructions to ensure successful experimental results, along with notes on troubleshooting and how to avoid pitfalls. Valuable appendices summarize changes in mass arising from protein modification, as well as consensus sequences for protein modification.
Comprehensive and up-to-date, Protein Sequencing Protocols, Second Edition, provides both novice and expert investigators alike a ready source of easy-to-follow protocols that simplify choosing the most appropriate method for protein sequence determination and thus help assure robust results.