E-Book Overview
John R. Chapman recreates the success of his earlier acclaimed book (Protein and Peptide Analysis by Mass Spectrometry, 1996) with a major new collection of cutting-edge methods in the analysis of proteins and peptides. Contributed by well-established investigators, each chapter provides background information before guiding the researcher step-by-step through the experimental process, with detailed instructions to help assure experimental success. The applications covered range widely and include protein sequencing, higher-order structure determination, epitope mapping, kinetics, quantitation, glycosylation analysis, and bacterial typing. Authoritative and eminently practical, Mass Spectrometry of Proteins and Peptides demonstrates the full scope of this versatile analytical technique. All protein chemists and biochemists, as well as bioanalytical scientists, who want to exploit these powerful methods in their work, will find this book indispensable.
E-Book Content
Methods in Molecular Biology
TM
VOLUME 146
Mass Spectrometry of Proteins and Peptides Edited by
John R. Chapman
HUMANA PRESS
Peptide Sequencing by Tandem MS
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1 De Novo Peptide Sequencing by Nanoelectrospray Tandem Mass Spectrometry Using Triple Quadrupole and Quadrupole/Time-of-Flight Instruments Andrej Shevchenko, Igor Chernushevich, Matthias Wilm, and Matthias Mann 1. Introduction Recent developments in technology and instrumentation have made mass spectrometry the method of choice for the identification of gel-separated proteins using rapidly growing sequence databases (1). Proteins with a full-length sequence present in a database can be identified with high certainty and high throughput using the accurate masses obtained by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry peptide mapping (2). Simple protein mixtur