Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells
Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells
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DOI：
10.1074/mcp.m900203-mcp200
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发表时间：
2009-10-01
影响因子：
7
通讯作者：
Marin-Castano, Maria E.
中科院分区：
文献类型：
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作者：
Alcazar, Oscar;Hawkridge, Adam M.;Marin-Castano, Maria E.
视网膜相关性黄斑变性（AMD）是工业化世界中老年人口中法律的失明的主要原因，仅在美国就影响约1400万人。吸烟是AMD的主要环境危险因素，而对苯二酚是香烟烟雾中的主要成分。对苯二酚可诱导人视网膜色素上皮细胞膜泡的形成。水泡可能积累并最终首先导致RPE下沉积物，然后形成玻璃疣，这是患有AMD的眼睛中的突出组织病理学特征。为了更好地了解早期AMD的发病机制，我们试图研究RPE滤过泡的蛋白质组学特征。对分离的泡进行SDS-PAGE分级分离，并通过LCMS/MS分析凝胶内胰蛋白酶消化的肽，从而鉴定出总共314种蛋白质。鉴定的蛋白质主要参与氧化磷酸化、细胞连接、粘着斑、细胞骨架调节和免疫原性过程。重要的是，basigin和基质金属蛋白酶-14，参与细胞外基质重塑的关键蛋白质，在RPE滤过泡中被鉴定，并且在AMD患者中更普遍。总之，我们的研究结果表明，第一次，在眼睛疾病的RPE泡的潜在参与，并阐明了人类病理学中的细胞衍生的微泡的含义。Molecular & Cellular Proteomics 8：2201-2211，2009.
Age-related macular degeneration (AMD) is the leading cause of legal blindness among the elderly population in the industrialized world, affecting about 14 million people in the United States alone. Smoking is a major environmental risk factor for AMD, and hydroquinone is a major component in cigarette smoke. Hydroquinone induces the formation of cell membrane blebs in human retinal pigment epithelium (RPE). Blebs may accumulate and eventually contribute first to sub-RPE deposits and then drusen formation, which is a prominent histopathologic feature in eyes with AMD. As an attempt to better understand the mechanisms involved in early AMD, we sought to investigate the proteomic profile of RPE blebs. Isolated blebs were subjected to SDS-PAGE fractionation, and in-gel trypsin-digested peptides were analyzed by LCMS/MS that lead to the identification of a total of 314 proteins. Identified proteins were predominantly involved in oxidative phosphorylation, cell junction, focal adhesion, cytoskeleton regulation, and immunogenic processes. Importantly basigin and matrix metalloproteinase-14, key proteins involved in extracellular matrix remodeling, were identified in RPE blebs and shown to be more prevalent in AMD patients. Altogether our findings suggest, for the first time, the potential involvement of RPE blebs in eye disease and shed light on the implication of cell-derived microvesicles in human pathology. Molecular & Cellular Proteomics 8: 2201-2211, 2009.