Mass Spectrometry of Proteins and Peptides

When the last edition of this book was published in 2000, the field of proteomics was in its infancy. At that time, multidimensional liquid chromatographic separations were being introduced as an alternative to traditional gel-based techniques for separating complex protein and peptide mixtures prior to mass spectrometric detection. Today, this approach referred to as shotgun proteomics is considered routine for lar- scale global analyses of protein mixtures. Now in its adolescence, proteomics is fundamentally transforming biological and medical research. Much of this transformation can be attributed to technological advancements, particularly in mass spectrometry. Much wider accessibility of hi- resolution and mass measurement accuracy instrumentation in recent years has ini- ated a new revolution in the field by providing more reliable data and shifting the focus from cataloging proteins to precisely quantifying changes in protein abundance over time and in response to stimuli. Advanced mass spectrometers and novel ion d- sociation schemes such as electron transfer/capture dissociation make it possible to venture boldly into the maze of protein posttranslational modifications, which are an integral component of understanding functional proteomics in the spatial and t- poral domains. Another area that has benefited from these advancements is top-down proteomics, an emerging method essential for characterizing various protein variants that has potentially high impact in biomedical research.

Comprehensive overview of latest mass spectrometric techniques for protein analysis Step-by-step methods complete with detailed instructions for successful implementation Written by established experts with hands-on experience Extensive, up-to-date references arranged by application Includes supplementary material: sn.pub/extras

Inhalt
Generation and Analysis of Multidimensional Protein Identification Technology Datasets.- Quantitative Peptide and Protein Profiling by Mass Spectrometry.- Label-Free Relative Quantitation of Prokaryotic Proteomes Using the Accurate Mass and Time Tag Approach.- Classical Proteomics: Two-Dimensional Electrophoresis/MALDI Mass Spectrometry.- The Use of Difference In-Gel Electrophoresis for Quantitation of Protein Expression.- Liquid-Chromatography-Mass Spectrometry of Thylakoid Membrane Proteins.- High Accuracy Mass Spectrometry in Large-Scale Analysis of Protein Phosphorylation.- Manual Validation of Peptide Sequence and Sites of Tyrosine Phosphorylation from MS/MS Spectra.- Assigning Glycosylation Sites and Microheterogeneities in Glycoproteins by Liquid Chromatography/Tandem Mass Spectrometry.- Structure Analysis of N -Glycoproteins.- Capillary Zone Electrophoresis-Mass Spectrometry for the Characterization of Isoforms of Intact Glycoproteins.- Top-Down Proteomics on a High-field Fourier Transform Ion Cyclotron Resonance Mass Spectrometer.- Capillary Isoelectric Focusing/Reversed Phase Liquid Chromatography/Mass Spectrometry.- Integrating Accelerated Tryptic Digestion into Proteomics Workflows.- Hydrogen/Deuterium Exchange Mass Spectrometry.- Mass Spectrometry Detection and Characterization of Noncovalent Protein Complexes.- Chemical Cross-Linking for ProteinProtein Interaction Studies.- Tissue Analysis with High-Resolution Imaging Mass Spectrometry.- Proteomic Global Profiling for Cancer Biomarker Discovery.- Analysis of Protein Glycosylation and Phosphorylation Using Liquid Phase Separation, Protein Microarray Technology, and Mass Spectrometry.- Transthyretin Mass Determination for Detection of Transthyretin Familial Amyloid.- Characterization of Microorganisms byMALDI Mass Spectrometry.- Mass Spectrometric Characterization of Neuropeptides.- Peptide and Protein Ion/Ion Reactions in Electrodynamic Ion Traps: Tools and Methods.- Electron Capture Dissociation LC/MS/MS for Bottom-Up Proteomics.- Two-Dimensional Ion Mobility Analyses of Proteins and Peptides.- Proteomics for Validation of Automated Gene Model Predictions.- Support Vector Machines for Improved Peptide Identification from Tandem Mass Spectrometry Database Search.