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Global differential non-gel proteomics by quantitative and stable labeling of tryptic peptides with oxygen-18

An Staes (UGent) , Hans Demol (UGent) , Jozef Van Damme (UGent) , Lennart Martens (UGent) , Joël Vandekerckhove (UGent) and Kris Gevaert (UGent)
(2004) JOURNAL OF PROTEOME RESEARCH. 3(4). p.786-791
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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
TRYPSIN, nongel proteomics, QUANTIFICATION, IDENTIFICATION, mass spectrometry, oxygen-18, differential analysis, COFRADIC, DESORPTION/IONIZATION MASS-SPECTROMETRY, CATALYZED O-16-TO-O-18 EXCHANGE, PROTEIN EXPRESSION, INTERNAL STANDARDS, AFFINITY TAGS, MIXTURES, PHOSPHORYLATION

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Chicago
Staes, An, Hans Demol, Jozef Van Damme, Lennart Martens, Joël Vandekerckhove, and Kris Gevaert. 2004. “Global Differential Non-gel Proteomics by Quantitative and Stable Labeling of Tryptic Peptides with Oxygen-18.” Journal of Proteome Research 3 (4): 786–791.
APA
Staes, A., Demol, H., Van Damme, J., Martens, L., Vandekerckhove, J., & Gevaert, K. (2004). Global differential non-gel proteomics by quantitative and stable labeling of tryptic peptides with oxygen-18. JOURNAL OF PROTEOME RESEARCH, 3(4), 786–791.
Vancouver
1.
Staes A, Demol H, Van Damme J, Martens L, Vandekerckhove J, Gevaert K. Global differential non-gel proteomics by quantitative and stable labeling of tryptic peptides with oxygen-18. JOURNAL OF PROTEOME RESEARCH. 2004;3(4):786–91.
MLA
Staes, An, Hans Demol, Jozef Van Damme, et al. “Global Differential Non-gel Proteomics by Quantitative and Stable Labeling of Tryptic Peptides with Oxygen-18.” JOURNAL OF PROTEOME RESEARCH 3.4 (2004): 786–791. Print.
@article{299851,
  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.},
  author       = {Staes, An and Demol, Hans and Van Damme, Jozef and Martens, Lennart and Vandekerckhove, Jo{\"e}l and Gevaert, Kris},
  issn         = {1535-3893},
  journal      = {JOURNAL OF PROTEOME RESEARCH},
  keyword      = {TRYPSIN,nongel proteomics,QUANTIFICATION,IDENTIFICATION,mass spectrometry,oxygen-18,differential analysis,COFRADIC,DESORPTION/IONIZATION MASS-SPECTROMETRY,CATALYZED O-16-TO-O-18 EXCHANGE,PROTEIN EXPRESSION,INTERNAL STANDARDS,AFFINITY TAGS,MIXTURES,PHOSPHORYLATION},
  language     = {eng},
  number       = {4},
  pages        = {786--791},
  title        = {Global differential non-gel proteomics by quantitative and stable labeling of tryptic peptides with oxygen-18},
  url          = {http://dx.doi.org/10.1021/pr049956p},
  volume       = {3},
  year         = {2004},
}

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