期刊
JOURNAL OF PROTEOMICS
卷 75, 期 3, 页码 857-867出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2011.09.027
关键词
Cysteinyl peptide enrichment; iTRAQ reagents; Proteome coverage; Amniotic fluid proteome
资金
- Ministry of Health, Czech Republic [NS/10382-3/2009]
- Czech Science Foundation GACR [301/09/P241]
Shotgun proteomics is capable of characterizing differences in both protein quality and quantity, and has been applied in various biomedical applications. Unfortunately, the high complexity and dynamic range of proteins in studied samples, clinical in particular, often hinders the identification of relevant proteins. Indeed, information-rich, low abundance proteins often remain undetected, whereas repeatedly reported altered concentrations in high abundance proteins are often ambiguous and insignificant. Several techniques have therefore been developed to overcome this obstacle and provide a deeper insight into the proteome. Here we report a novel approach, which enables iTRAQ reagent quantitation of peptides fractionated based on presence of a cysteine residue (thus CysTRAQ). For the first time, we prove that iTRAQ quantitation is fully compatible with cysteinyl peptide enrichment and is not influenced by the fractionation process. Moreover, the employment of the method combined with high-resolution TripleTOF 5600 mass spectrometer for very fast MS/MS acquisition in human amniotic fluid analysis significantly increased the number of identified proteins, which were simultaneously quantified owing to the introduction of iTRAQ labeling. We herein show that CysTRAQ is a robust and straightforward method with potential application in quantitative proteomics experiments, i.e. as an alternative to the ICAT reagent approach. (C) 2011 Elsevier B.V. All rights reserved.
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