Uhl P B, Szober C M, Amann B, Alge-Priglinger C, Ueffing M, Hauck S M, Deeg C A
Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University, Munich, Germany.
Department of Ophthalmology, Ludwig-Maximilians-University, Munich, Germany.
J Proteomics. 2014 Sep 23;109:50-62. doi: 10.1016/j.jprot.2014.06.020. Epub 2014 Jul 2.
Retinal pigment epithelium (RPE) builds the outer blood-retinal barrier of the eye and plays an important role in pathogenesis of the sight threatening disease equine recurrent uveitis (ERU). ERU is a spontaneous autoimmune mediated inflammatory disease characterised by the breakdown of the outer blood-retinal barrier and an influx of autoaggressive T-cells into the inner eye. Therefore, identification of molecular mechanisms contributing to changed function of blood-retinal barrier in ERU is important for the understanding of pathophysiology. Cell surface proteins of RPE collected from healthy horses and horses with ERU were captured by in situ biotinylation and analysed with high resolution mass spectrometry coupled to liquid chromatography (LC-MS/MS) to identify differentially expressed proteins. With label free differential proteomics, a total of 27 differently expressed cell surface proteins in diseased RPE could be detected. Significant down-regulation of three very interesting proteins, synaptotagmin 1, basigin and collectrin was verified and further characterised.
We applied an innovative and successful method to detect changes in the plasma cell surface proteome of RPE cells in a spontaneous inflammatory eye disease, serving as a valuable model for human autoimmune uveitis. We were able to identify 27 differentially expressed plasma cell membrane proteins, including synaptotagmin 1, basigin and collectrin, which play important roles in cell adhesion, transport and cell communication.
视网膜色素上皮（RPE）构成眼睛的外血视网膜屏障，在威胁视力的疾病马反复性葡萄膜炎（ERU）的发病机制中起重要作用。ERU是一种自发性自身免疫介导的炎症性疾病，其特征是外血视网膜屏障破坏以及自身攻击性T细胞流入眼内。因此，确定导致ERU中外血视网膜屏障功能改变的分子机制对于理解病理生理学很重要。通过原位生物素化捕获从健康马和患有ERU的马收集的RPE细胞表面蛋白，并用液相色谱联用的高分辨率质谱（LC-MS/MS）进行分析，以鉴定差异表达的蛋白质。通过无标记差异蛋白质组学，在患病的RPE中总共检测到27种差异表达的细胞表面蛋白。验证并进一步表征了三种非常有趣的蛋白，即突触结合蛋白1、基底膜蛋白和collectrin的显著下调。
我们应用了一种创新且成功的方法来检测自发性炎症性眼病中RPE细胞浆膜蛋白质组的变化，该疾病可作为人类自身免疫性葡萄膜炎的有价值模型。我们能够鉴定出27种差异表达的浆细胞膜蛋白，包括突触结合蛋白1、基底膜蛋白和collectrin，它们在细胞粘附、运输和细胞通讯中起重要作用。
