Proteome Analysis of Human Aqueous Humor
Proteome Analysis of Human Aqueous Humor
复制标题
DOI：
10.1167/iovs.10-5531
复制
发表时间：
2010-10-01
影响因子：
4.4
通讯作者：
Fautsch, Michael P.
中科院分区：
文献类型：
--
作者：
Chowdhury, Uttio Roy;Madden, Benjamin J.;Fautsch, Michael P.
目的.人的房水（hAH）在眼睛的前房内提供营养和免疫。hAH的蛋白质组成的表征将鉴定参与维持眼前节组织的稳态环境的分子。本研究对hAH的蛋白质组进行了分析。将在选择性白内障手术期间获得的hAH样品分成三个匹配组，并免疫耗竭白蛋白、IgG、伊加、单珠蛋白、抗胰蛋白酶和转铁蛋白。通过凝胶电泳分离每组的还原和变性蛋白质（20 μ g）。从三个凝胶泳道中的每一个切下33个凝胶切片（n = 99），用胰蛋白酶消化，并进行纳流液相色谱电喷雾电离串联质谱（nano-LC-ESI-MS/MS）。还通过基于抗体的蛋白质阵列分析了hAH的蛋白质组分，并对选择的蛋白质进行定量。在hAH中共鉴定出676个蛋白质。在通过nano-LC-ESI-MS/MS鉴定的355种蛋白质中，在所有三个组中发现了206种。通过nano-LC-ESI-MS/MS鉴定的大多数蛋白质具有催化、酶促和结构性质。使用基于抗体的蛋白质阵列，鉴定了328种细胞因子、趋化因子和受体。大多数定量蛋白质的浓度范围为0.1 - 2.5 ng/mL。通过nano-LC-ESIMS/MS和抗体蛋白芯片鉴定了10个蛋白质。hAH的蛋白质组学分析鉴定了676个非冗余蛋白。其中80%以上的蛋白质是新鉴定的。该蛋白质组的阐明将为蛋白质功能分析和鉴定与眼前节疾病相关的差异表达标志物奠定基础。（Invest Ophthalmol维斯科学。2010;51：4921-4931）DOI：10.1167/iovs.10-5531
PURPOSE. Human aqueous humor (hAH) provides nutrition and immunity within the anterior chamber of the eye. Characterization of the protein composition of hAH will identify molecules involved in maintaining a homeostatic environment for anterior segment tissues. The present study was conducted to analyze the proteome of hAH.METHODS. hAH samples obtained during elective cataract surgery were divided into three matched groups and immunodepleted of albumin, IgG, IgA, haploglobin, antitrypsin, and transferrin. Reduced and denatured proteins (20 mu g) from each group were separated by gel electrophoresis. Thirty-three gel slices were excised from each of three gel lanes (n = 99), digested with trypsin, and subjected to nanoflow liquid chromatography electrospray ionization tandem mass spectrometry (nano-LC-ESI-MS/MS). The protein component of hAH was also analyzed by antibody-based protein arrays, and selected proteins were quantified.RESULTS. A total of 676 proteins were identified in hAH. Of the 355 proteins identified by nano-LC-ESI-MS/MS, 206 were found in all three groups. Most of the proteins identified by nano-LC-ESI-MS/MS had catalytic, enzymatic, and structural properties. Using antibody-based protein arrays, 328 cytokines, chemokines, and receptors were identified. Most of the quantified proteins had concentrations that ranged between 0.1 and 2.5 ng/mL. Ten proteins were identified by both nano-LC-ESIMS/MS and antibody protein arrays.CONCLUSIONS. Proteomic analysis of hAH identified 676 nonredundant proteins. More than 80% of these proteins are novel identifications. The elucidation of the aqueous proteome will establish a foundation for protein function analysis and identification of differentially expressed markers associated with diseases of the anterior segment. (Invest Ophthalmol Vis Sci. 2010;51:4921-4931) DOI:10.1167/iovs.10-5531