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Profound Re-Organization of Cell Surface Proteome in Equine Retinal Pigment Epithelial Cells in Response to In Vitro Culturing

التفاصيل البيبلوغرافية
العنوان: Profound Re-Organization of Cell Surface Proteome in Equine Retinal Pigment Epithelial Cells in Response to In Vitro Culturing
المؤلفون: Marius Ueffing, Cornelia A. Deeg, Claudia Alge-Priglinger, Kerstin N. Euler, Christoph M. Szober, Kristina J. H. Fröhlich, Stefanie M. Hauck
المصدر: International Journal of Molecular Sciences, Vol 13, Iss 11, Pp 14053-14072 (2012)
بيانات النشر: MDPI AG, 2012.
سنة النشر: 2012
المجموعة: LCC:Biology (General)
LCC:Chemistry
مصطلحات موضوعية: equine recurrent uveitis, membrane protein, outer blood-retinal barrier, retinal pigment epithelium cells, cell line, proteomics, Biology (General), QH301-705.5, Chemistry, QD1-999
الوصف: The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses’ vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, analyzed by high resolution mass spectrometry coupled to liquid chromatography (LC MS/MS), and the most interesting candidates were validated by PCR, immunoblotting and immunocytochemistry. A total of 112 proteins were identified, of which 84% were cell surface membrane proteins. Twenty-three of these proteins were concurrently expressed by both cell states, 28 proteins exclusively by native RPE cells. Among the latter were two RPE markers with highly specialized RPE functions: cellular retinaldehyde-binding protein (CRALBP) and retinal pigment epithelium-specific protein 65kDa (RPE65). Furthermore, 61 proteins were only expressed by cultured RPE cells and absent in native cells. As we believe that initiating events, leading to the breakdown of the outer blood-retinal barrier, take place at the cell surface of RPE cells as a particularly exposed barrier structure, this differential characterization of cell surface proteomes of native and cultured equine RPE cells is a prerequisite for future studies.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1422-0067
Relation: http://www.mdpi.com/1422-0067/13/11/14053; https://doaj.org/toc/1422-0067
DOI: 10.3390/ijms131114053
URL الوصول: https://doaj.org/article/4ddbc62581994c33a0cae31cf34eea42
رقم الأكسشن: edsdoj.4ddbc62581994c33a0cae31cf34eea42
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:14220067
DOI:10.3390/ijms131114053