Global Differential Non-Gel Proteomics by Quantitative and Stable Labeling of Tryptic Peptides with Oxygen-18
- 7 May 2004
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of Proteome Research
- Vol. 3 (4), 786-791
- https://doi.org/10.1021/pr049956p
Abstract
We describe a protocol for quantitative labeling of tryptic peptides with oxygen-18. Proteins are first digested in natural water with trypsin, the pH is then lowered to 4.5 and the mixture is dried. Oxygen-18 water is added and two oxygen-18 atoms are incorporated at the peptides' carboxyl termini. Trypsin is finally inactivated by cysteine alkylation under denaturing conditions, which blocks oxygen back-exchange. The general value of this labeling strategy for differential proteomics is illustrated by the analysis and identification of several couples of differently labeled amino terminal peptides isolated from a human platelet proteome by a previously described chromatographic procedure. Keywords: nongel proteomics • mass spectrometry • oxygen-18 • differential analysis • COFRADICKeywords
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