Proteome profiling of corneal epithelium and identification of marker proteins for keratoconus, a pilot study.
Proteome profiling of corneal epithelium and identification of marker proteins for keratoconus, a pilot study.
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DOI：
10.1016/j.exer.2005.06.009
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发表时间：
2006-02-01
影响因子：
3.4
通讯作者：
Orntoft, TF
中科院分区：
文献类型：
--
作者：
Nielsen, K;Vorum, H;Orntoft, TF
本研究的目的是鉴定圆锥角膜和正常角膜上皮样本之间差异表达的角膜蛋白质。从6例圆锥角膜和6例近视患者（对照）的角膜上皮中分离蛋白，并通过二维凝胶电泳分离。使用6%和12% SDS-PAGE凝胶分离低分子量和高分子量蛋白质。将凝胶进行银染色，并通过Melanie II软件定义蛋白质斑点。提取与圆锥角膜和对照相比表达变化最大的蛋白质，用胰蛋白酶消化，并通过质谱法鉴定。在每次获得中检测到大约200-500个蛋白质点。19个点被鉴定为在圆锥角膜和参考上皮之间差异表达，包括细胞角蛋白3（< 7.8倍）、凝溶胶蛋白（1.6倍）、S100 A4（1.9倍）和烯醇化酶1（0.72倍）。另一种被鉴定为高水平的蛋白质是细胞角蛋白12。凝溶胶蛋白、细胞角蛋白3和细胞角蛋白12先前已被描述为参与其他角膜疾病。三种蛋白质，凝溶胶蛋白，α烯醇化酶，和S100 A4被确定为在圆锥角膜相比，参考上皮细胞的差异表达，因此可能参与发病机制。(c)2005爱思唯尔有限公司保留所有权利。
The purpose of this study is to identify corneal proteins differentially expressed between keratoconus and normal epithelial samples. Proteins from the corneal epithelium were isolated from 6 keratoconus and 6 myopia patients (controls) and separated by 2D-gel electrophoresis. Six % and 12% SDS-PAGE gels were used to separate low and high molecular weight proteins. Gels were silver stained and protein spots were defined by Melanie II software. The proteins that were most altered in expression comparing keratoconus and controls were extracted, trypsin-digested, and identified by mass spectroscopy. Approximately 200-500 protein spots were detected on each get. Nineteen spots were identified as differentially expressed between keratoconus and reference epithelium including cytokeratin 3 ( < 7.8 fold), gelsolin (1.6 fold), S100A4 (1.9 fold), and enolase 1 (0.72 fold). Another identified protein found at very high levels was cytokeratin 12. Gelsolin, cytokeratin 3, and cytokeratin 12 have previously been described to be involved in other corneal diseases. Three proteins, gelsolin, alpha enolase, and S100A4 were identified to be differentially expressed in keratoconus compared to reference epithelium and thus may be involved in the pathogenesis. (c) 2005 Elsevier Ltd. All rights reserved.