Journal
JOURNAL OF PROTEOMICS
Volume 74, Issue 2, Pages 151-166Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2010.10.002
Keywords
Aqueous humour; iTRAQ; (R) Orbitrap; 2D shotgun; Eye proteomics; 1D gel
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Funding
- Austrian Academy of Sciences
- Austrian Federal Ministry for Science and Research
- Austrian Science Fund FWF
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In this study, we report a comparative and quantitative analysis by mass spectrometry of the protein content of aqueous humour from cataract (control) patients. In addition to protein profiling, the approach is layered with quantitative proteomics using the iTRAQ (R) methodology. Aqueous humour from ten clinically-matched patients was collected and depleted of albumin and immunoglobulin G. Pairs of patient material were pooled and divided into three aliquots for subsequent analysis by alternative proteomic approaches. Excluding keratin, trypsin, residual albumin and immunoglobulins, a total of 198 protein groups were identified across the entire study. Relative protein quantitation with iTRAQ (R) revealed that 88% of the proteins had a maximal +/-2-fold differential regulation between 3 of the 4 labelled samples, indicating minimal variation. The identified proteins were categorised by gene ontology and one third of the proteins were annotated as extracellular. The major molecular functions of the proteins in aqueous humour are binding (protein, metal ion, heparin, and DNA) and inhibition of proteolytic activity. Complementary to molecular function, the predominant biological processes for the proteins in aqueous humour are assigned to inflammatory and immune responses, and transport. (C) 2010 Elsevier B.V. All rights reserved.
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