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Protein Sequencing Protocols

Series: Methods in Molecular Biology

by Bryan John Smith

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Main description:

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in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches.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.

Contents:

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

PRODUCT DETAILS

ISBN-13: 9781617372841
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 506
Weight: 817g
Availability: Available
Subcategories: Biochemistry

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