文档介绍：1
PROTEOMICS: THE
BASIC OVERVIEW
Daniel Figeys
Department of Biochemistry, Microbiology,
and Immunology, University of Ottawa, Ottawa, Canada
Introduction 2
Deoxyribonucleic Acid to Ribonucleic Acid 4
Ribonucleic Acid to Protein 6
Proteome Handling in Classical and Functional Proteomics 6
Protein Purification 6
Cell Culture 8
Other Source of Samples 8
Subpopulation of Cells 10
ponents 11
Protein Separation by Two-Dimensional Gel Electrophoresis 12
Principles of Two-Dimensional Gel Electrophoretic Separation 13
Detection of Proteins Separated by Two-Dimensional Gel Electrophoresis 15
Software to Handle Two-Dimensional Gel Electropherogram 17
Potein Processing 17
Blot Digestion of Proteins 18
In-gel Digestion of Proteins 19
Protein Identification 19
Edman Degradation 19
Industrial Proteomics: Applications for Biotechnology and Pharmaceuticals, edited by Daniel Figeys
ISBN 0-471-45714-0 Copyright © 2005 by John Wiley & Sons, Inc.
1
2 PROTEOMICS: THE BASIC OVERVIEW
Protein Identification by Mass Spectrometry 20
Hybrid Instruments 26
Bioinformatics for the Identification of Proteins Based on MS/MS 46
Current Challenges in Proteomics 49
Classical Proteomics 49
Gel-Free Analysis: An Alternative Differential Display 50
Fourier Transofrm Mass Spectrometery for Gel-Free Analysis and
Single-Peptide-Based Identification 53
References 54
List of Abbreviations
2D Two dimensional
b-gal b-galactosidase
bp base pair
CZE capillary zone electrophoresis
ESI electrospray ionization
EST expressed sequence tag
FTMS Fourier transform mass spectrometer
HPLC high-performance liquid chromatography
. inner diameter
ICAT isotope-coded affinity tag
IPG immobilized pH gradient
MALDI matrix-assisted laser desorption ionization
MHC major plex
Micro-ESI microelectrospray ionization
MS/MS tandem mass spectrum
MS mass spectr(um/ometer)
MW molecular weight
Nano-ESI nanoelectrospray ionization
. outer