Zhang Pingbo, Kirby David, Dufresne Craig, Chen Yan, Turner Randi, Ferri Sara, Edward Deepak P, Van Eyk Jennifer E, Semba Richard D
Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Thermo Fisher Scientific, West Palm Beach, FL, USA.
Proteomics. 2016 Apr;16(7):1146-53. doi: 10.1002/pmic.201500188. Epub 2016 Mar 11.
The iris is a fine structure that controls the amount of light that enters the eye. The ciliary body controls the shape of the lens and produces aqueous humor. The retinal pigment epithelium and choroid (RPE/choroid) are essential in supporting the retina and absorbing light energy that enters the eye. Proteins were extracted from iris, ciliary body, and RPE/choroid tissues of eyes from five individuals and fractionated using SDS-PAGE. After in-gel digestion, peptides were analyzed using LC-MS/MS on an Orbitrap Elite mass spectrometer. In iris, ciliary body, and RPE/choroid, we identified 2959, 2867, and 2755 nonredundant proteins with peptide and protein false-positive rates of <0.1% and <1%, respectively. Forty-three unambiguous protein isoforms were identified in iris, ciliary body, and RPE/choroid. Four "missing proteins" were identified in ciliary body based on ≥2 proteotypic peptides. The mass spectrometric proteome database of the human iris, ciliary body, and RPE/choroid may serve as a valuable resource for future investigations of the eye in health and disease. The MS proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifiers PXD001424 and PXD002194.
虹膜是一种精细的结构,可控制进入眼睛的光量。睫状体控制晶状体的形状并产生房水。视网膜色素上皮和脉络膜(RPE/脉络膜)对于支持视网膜和吸收进入眼睛的光能至关重要。从五个人的眼睛的虹膜、睫状体和RPE/脉络膜组织中提取蛋白质,并使用SDS-PAGE进行分级分离。凝胶内消化后,使用Orbitrap Elite质谱仪通过LC-MS/MS分析肽段。在虹膜、睫状体和RPE/脉络膜中,我们分别鉴定出2959、2867和2755种非冗余蛋白质,肽段和蛋白质的假阳性率分别<0.1%和<1%。在虹膜、睫状体和RPE/脉络膜中鉴定出43种明确的蛋白质异构体。基于≥2个蛋白型肽段在睫状体中鉴定出4种“缺失蛋白质”。人类虹膜、睫状体和RPE/脉络膜的质谱蛋白质组数据库可能是未来健康和疾病状态下眼部研究的宝贵资源。质谱蛋白质组学数据已通过PRIDE合作伙伴库存入蛋白质组交换联盟,数据集标识符为PXD001424和PXD002194。