[분석] mass spectrometry를 이용한 phosphoprotein and phosphopeptide 분석
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소개글
프로테오믹스에서 사용하는 phospho-protein and -peptide 분석의 모든 방법
정말 잘 정리된 자료...
목차
-Overview of protein phosphorylation analysis by mass spectrometry
-Phosphorylation Facts
-The chemical structures of the phosphoamino acids
-Phosphopeptides are generally difficult to analyze by mass spectrometry (MS) for several reasons.
-Comparison of mass spectrometry-based technoques with other methods for phosphorylation analysis of peptides
-An overview of techniques for enrichment and analysis of phosphorylated proteins or peptidesusing mass spectrometry (MS)-based detection methods
-Sample Preparation
-Fractionation
-IMAC (Immobilized Metal Affinity Chromatography)
-Direct MS of phosphopeptides bound to IMAC beads
-Antibodies
-Chemical tagging of phosphorylation sites
-Chemical modification methods
-Recognition of phosphopeptides
-Post-source decay
-Positive ion MALDI TOF mass spectra
-Precusor ion scan
-Neutral loss scanning
-Stepped skimmer potential
-32P detection
-Identification of phosphorylation sites
-In-source decay
-Electron capture dissociation
-Quantitation of phosphorylation
-Peak intensities
-Stable isotope amino acid or 15N in vivo labeling
-Isotopic labeling
-ICAT: Isotope-Coded Affinity Tag
-ICAT: Isotope-Coded Affinity Tag
-Conclusions and future directions
-Additional Readings
본문내용
Phosphorylation Facts
Serine and threonine residues undergo phosphorylation more often than tyrosine residues. The phosphoamino acid content ratio (pSer:pThr:pTyr) of a vertebrate cell is 1800:200:1. There is a higher gain in signals involving tyrosine phosphorylation because it is less abundant and more tightly regulated.
One-third of all proteins in a eukaryotic cell are phosphorylated at any one time.
There are several commercially available antibodies that enrich for tyrosine phosphorylated proteins. Similar antibodies to enrich serine or threonine phosphorylated proteins are not currently available.
Phosphorylation is heterogeneous. Most phosphoproteins undergo phosphorylation on more than one residue
참고 자료
R. Aebersold and M. Mann, Mass spectrometry-based proteomics, Nature (2003), 422, 198-207.
M. B. Goshe and R. D. Smith, 밪table isotope-coded proteomic mass spectrometry.