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Proteomic analysis of human cataract aqueous humour: Comparison of one-dimensional gel LCMS with two-dimensional LCMS of unlabelled and iTRAQ®-labelled specimens
- Source :
- Journal of proteomics. 74(2)
- Publication Year :
- 2010
-
Abstract
- 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® 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® 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.
- Subjects :
- Proteomics
Chromatography
Chemistry
Aqueous humour
Quantitative proteomics
Biophysics
Albumin
Trypsin
Biochemistry
Cataract
Mass Spectrometry
Aqueous Humor
chemistry.chemical_compound
Isotope Labeling
medicine
Extracellular
Humans
Electrophoresis, Gel, Two-Dimensional
Electrophoresis, Polyacrylamide Gel
Eye Proteins
Quantitative analysis (chemistry)
DNA
medicine.drug
Chromatography, Liquid
Subjects
Details
- ISSN :
- 18767737
- Volume :
- 74
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of proteomics
- Accession number :
- edsair.doi.dedup.....893a0d7a2266f2fc53a8407959697bb4