Purchase this article with an account.
O. Alcazar, A. M. Hawkridge, D. C. Muddiman, S. K. Bhattacharya, M. E. Marin-Castano; Proteomic Characterization of Hydroquinone-Induced Blebs in a Human RPE Cell Line. Invest. Ophthalmol. Vis. Sci. 2008;49(13):484.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Hydroquinone (HQ), a major component of cigarette smoke, which is a risk factor in dry Age-related macular degeneration (AMD), induces formation of non-lethal RPE membrane bleb. This study aims to determine composition of HQ induced RPE bleb proteome, HQ induced bleb glycoproteome, and phosphoproteome in a human RPE cell line.
Human RPE cells were treated with 100 µM HQ for 6 h to allow bleb formation. Blebs were isolated and bleb proteins fractionated on 4-20% gradient gels were assayed for total glycosylation or phosphorylation (GelCode Glycoprotein and Phosphoprotein Staining kits, Pierce; and Pro-Q Diamond and Pro-Q Emerald, Molecular Probes) and subsequently subjected to liquid chromatography tandem mass spectrometry.
Mass spectrometry identified a total of 314 proteins in blebs including glycosylation enzymes, for example, protein O-fucosyltransferase 2. KEGG pathway analysis found identified proteins involved in oxidative phosphorylation, cell junction, and actin cytoskeleton regulation. Glycoprotein and phosphoprotein analyses revealed characteristic patterns in HQ-induced blebs. The blebs showed the presence of several glycoproteins such as phosphatidylinositol glycan class S and Leprecan.
Non-lethal blebs induced with HQ in RPE cells show a characteristic protein profile pattern. Further investigation of these proteins may be useful to identify potential biomarkers and provide insight into the mechanisms leading to dry AMD.
This PDF is available to Subscribers Only