Differential Proteomics Incorporating iTRAQ Labeling and Multi-dimensional Separations
- 4k Downloads
Considerable effort is currently being expended to integrate newly developed “omics”-based approaches (proteomics, transcriptomics, and metabonomics) into preclinical safety evaluation workflows in the hope that more sensitive prediction of toxicology can be achieved as reported by Waters and Fostel (Nat. Rev. Genet. 5(12):936–948, 2004) and Craig et al. (J. Proteome Res. 5(7):1586–1601, 2006). Proteomic approaches are well placed to contribute to this effort as (a) proteins are the metabolically active products of genes and, as such, may provide more sensitive and direct predictive information on drug-induced liabilities and (b) they have the potential to determine tissue leakage markers in peripheral fluids. Here, we describe a workflow for proteomic semi-quantitative expression profiling of liver from rats treated with a known hepatotoxicant using a multiplexed isobaric labeling strategy and multi-dimensional liquid chromatography.
Key wordsToxicoproteomics iTRAQ Proteomics Toxicology Liver
We would like to thank all the members of the PredTox Consortium. Funding is acknowledged under the EU FP6 Integrated Project, InnoMed. The UCD Conway Institute and the Proteome Research Centre is funded by the Programme for Research in Third Level Institutions (PRTLI), as administered by the Higher Education Authority (HEA) of Ireland.
- 8.iTRAQ Reagents Chemistry Reference Guide. (2004) [http://www3.appliedbiosystems.com/cms/groups/psm_marketing/documents/generaldocuments/cms_041463.pdf]
- 15.Keller, A., et al. (2005) A uniform proteomics MS/MS analysis platform utilizing open XML file formats. Mol. Syst. Biol. Article number: 1, 2005.0017. pages 1–8.Google Scholar