蛋白质组学

蛋白质组学指南-来自杜克大学(Molecular Biologist’s Guide to Proteomics)

对蛋白组学方面的新手很有用。

MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, Mar. 2002, p. 39–63

Molecular Biologist’s Guide to Proteomics
Paul R. Graves1 and Timothy A. J. Haystead1,2*
Department of Pharmacology and Cancer Biology, Duke University,1 and Serenex Inc.,2 Durham, North Carolina 27710
INTRODUCTION........................................................................................................................................................40
Definitions.................................................................................................................................................................40
Proteomics Origins ..................................................................................................................................................40
Genome Information ...............................................................................................................................................41
Why Proteomics?......................................................................................................................................................41
Annotation of the genome...................................................................................................................................41
Protein expression studies..................................................................................................................................41
Protein function ...................................................................................................................................................42
Protein modifications ..........................................................................................................................................42
Protein localization and compartmentalization ...............................................................................................42
Protein-protein interactions ...............................................................................................................................42
Types of Proteomics.................................................................................................................................................42
Protein expression proteomics ............................................................................................................................42
Structural proteomics..........................................................................................................................................42
Functional proteomics.........................................................................................................................................42
TECHNOLOGY OF PROTEOMICS........................................................................................................................43
Separation and Isolation of Proteins.....................................................................................................................43
One- and two-dimensional gel electrophoresis .................................................................................................43
Alternatives to electrophoresis...........................................................................................................................44
Acquisition of Protein Structure Information ......................................................................................................45
Edman sequencing...............................................................................................................................................45
Mass spectrometry...............................................................................................................................................45
(i) Sample preparation....................................................................................................................................45
(ii) Sample ionization......................................................................................................................................45
(iii) Mass analysis ...........................................................................................................................................46
(iv) Types of mass spectrometers ...................................................................................................................48
(v) Peptide fragmentation...............................................................................................................................48
(vi) Our approach to mass spectrometry ......................................................................................................49
Database Utilization ................................................................................................................................................50
Peptide mass fingerprinting database searching .............................................................................................50
Amino acid sequence database searching .........................................................................................................50
De novo peptide sequence information..............................................................................................................52
Uninterpreted MS/MS data searching...............................................................................................................52
PROTEOMICS APPLICATIONS..............................................................................................................................53
Characterization of Protein Complexes.................................................................................................................53
Protein Expression Profiling ..................................................................................................................................53
Expression profiling by two-dimensional electrophoresis ...............................................................................53
Isotope-coded affinity tags ..................................................................................................................................53
Protein arrays.......................................................................................................................................................55
Proteomics Approach to Protein Phosphorylation...............................................................................................55
Phosphoprotein enrichment ...............................................................................................................................55
Phosphorylation site determination by Edman degradation ..........................................................................55
Phosphorylation site determination by mass spectrometry ............................................................................57
(i) Phosphopeptide sequencing by MS/MS ...................................................................................................57
(ii) Analysis of phosphopeptides by MALDI-TOF.......................................................................................58
Yeast and Proteomics............................................................................................................................58
Proteome Mining .....................................................................................................................................................58
Challenges for Proteomics......................................................................................................................................60
ACKNOWLEDGMENTS ............................................................................................................................................60
REFERENCES .............................................................................................................................................................60

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