May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Proteomic Characterization of Hydroquinone-Induced Blebs in a Human RPE Cell Line
Author Affiliations & Notes
  • O. Alcazar
    Ophthalmology, Bascom Palmer Eye Institute, Miami, Florida
  • A. M. Hawkridge
    Chemistry, North Carolina State University, Raleigh, North Carolina
  • D. C. Muddiman
    Chemistry, North Carolina State University, Raleigh, North Carolina
  • S. K. Bhattacharya
    Ophthalmology, Bascom Palmer Eye Institute, Miami, Florida
  • M. E. Marin-Castano
    Ophthalmology, Bascom Palmer Eye Institute, Miami, Florida
  • Footnotes
    Commercial Relationships  O. Alcazar, None; A.M. Hawkridge, None; D.C. Muddiman, None; S.K. Bhattacharya, None; M.E. Marin-Castano, None.
  • Footnotes
    Support  R01 EY015249-04
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 484. doi:https://doi.org/
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      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. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: proteomics • retinal pigment epithelium 
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