Improved Method for Differential Expression Proteomics Using Trypsin-catalyzed 18O Labeling with a Correction for Labeling Efficiency
نویسندگان
چکیده
منابع مشابه
Proteolytic 18O labeling for comparative proteomics: evaluation of endoprotease Glu-C as the catalytic agent.
Recently, proteolytic 18O labeling has been demonstrated as a promising strategy for comparative proteomic studies (Yao, X.; Freas, A.; Ramirez, J.; Demirev, P. A.; Fenselau, C. Anal. Chem. 2001, 73, 2836-42). In this approach, protein mixtures are digested in parallel in H216O and H218O and the ratios of isotopically distinct peptide products are measured by mass spectrometry. In the initial r...
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An enzymatic approach to label peptide N-termini with isotope-coded affinity tags is presented. This method exploits the high activity of trypsin for peptide synthesis in organic solvents. A cosubstrate containing a stable isotope-coded Arg residue and a biotin tag was synthesized. When the cosubstrate was incubated with tryptic peptides and trypsin in ethanol solution, the stable isotope-coded...
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Typical mass spectrometry-based protein lists from purified fractions are confounded by the absence of tools for evaluating contaminants. In this report, we compare the results of a standard survey experiment using an ion trap mass spectrometer with those obtained using dual isotope labeling and a Q-TOF mass spectrometer to quantify the degree of enrichment of proteins in purified subcellular f...
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ژورنال
عنوان ژورنال: Molecular & Cellular Proteomics
سال: 2007
ISSN: 1535-9476
DOI: 10.1074/mcp.t600029-mcp200