Abstract
Stable isotope labeling based on relative peptide/protein abundance measurements is commonly applied for quantitative proteomics. Recently, trypsin-catalyzed oxygen-18 labeling has grown in popularity due to its simplicity, cost-effectiveness, and its ability to universally label peptides with high sample recovery. In 18O labeling, both C-terminal carboxyl group atoms of tryptic peptides can be enzymatically exchanged with 18O, thus providing the labeled peptide with a 4 Da mass shift from the 16O-labeled sample. Peptide 18O labeling is ideally suited for generating a labeled “universal” reference sample used for obtaining accurate and reproducible quantitative measurements across large number of samples in quantitative discovery proteomics.
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Abbreviations
- ABC:
-
ammonium bicarbonate
- AMT:
-
accurate mass and time
- BCA:
-
bicinchoninic acid
- DTT:
-
dl-dithiothreitol
- ESI:
-
electrospray ionization
- FTICR:
-
Fourier transform ion cyclotron resonance
- IAA:
-
iodoacetamide
- I.D.:
-
inner diameter
- LC:
-
liquid chromatography
- MS:
-
mass spectrometry
- NET:
-
normalized elution time
- SPE:
-
solid phase extraction
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Acknowledgments
The authors thank NIH National Center for Research Resources (RR018522) for support of portions of this research and the Environmental Molecular Sciences Laboratory (EMSL) at Pacific Northwest National Laboratory (PNNL), a national scientific user facility sponsored by the US Department of Energy’s (DOE) Office of Biological and Environmental Research, for the development and use of the instrumentation applied in this research. PNNL is a multi-program national laboratory operated by Battelle Memorial Institute for the DOE under Contract DE-AC05-76RL01830.
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Qian, WJ., Petritis, B.O., Nicora, C.D., Smith, R.D. (2011). Trypsin-Catalyzed Oxygen-18 Labeling for Quantitative Proteomics. In: Gevaert, K., Vandekerckhove, J. (eds) Gel-Free Proteomics. Methods in Molecular Biology, vol 753. Humana Press. https://doi.org/10.1007/978-1-61779-148-2_3
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DOI: https://doi.org/10.1007/978-1-61779-148-2_3
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