Proteomic analysis of aqueous humour from patients with acute corneal rejection
Proteomic analysis of aqueous humour from patients with acute corneal rejection
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DOI：
10.1111/j.1600-0420.2005.00381.x
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发表时间：
2005-02-01
期刊：
影响因子：
--
通讯作者：
Ehlers, N
中科院分区：
文献类型：
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作者：
Funding, M;Vorum, H;Ehlers, N
目的：比较角膜排斥反应患者（患者）与白内障患者（对照组）的房水（AH）的基本蛋白质组组成。方法：采用Bradford方法分析了房水的总蛋白浓度，并采用二维（2D）凝胶电泳分析了蛋白质组成。图像分析用于检测2D凝胶中的蛋白质点，与对照组相比，患者中的蛋白质点增加了2倍以上。免疫印迹和质谱法确定了增加的斑点。结果：从患者的房水中含有显着较高的总蛋白浓度比AH从控制。在患者的2D凝胶中，共有31个斑点显著增加。斑点来源于白蛋白、α 1-抗胰蛋白酶、载脂蛋白J、细胞角蛋白II型、丝氨酸蛋白酶抑制剂和甲状腺素运载蛋白。总蛋白浓度校正后的斑点体积，来自白蛋白，细胞角蛋白II型和α 1-抗胰蛋白酶的10个斑点仍然显着increasing.Conclusion：AH的蛋白质组组成患者和对照组之间的显着差异。确定的蛋白质表明，AH的变化是由于至少三种不同的机制：水-血屏障的破坏，酶降解和释放局部合成的蛋白质。
Purpose: To compare the basic proteomic composition of aqueous humour (AH) from patients with corneal rejection (patients) with AH from patients with cataract (controls).Methods: Aqueous humour was analysed for total protein concentration using Bradford's method and for protein composition using two-dimensional (2D) gel electrophoresis. Image analysis was used to detect protein spots in 2D gels that were increased by more than factor 2 in patients as compared with controls. Increased spots were identified by immunoblotting and mass spectrometry.Results: Aqueous humour from patients contained significantly higher total protein concentration than did AH from controls. A total of 31 spots were significantly increased in 2D gels from patients. The spots were derived from albumin, alpha1-antitrypsin, apolipoprotein J, cytokeratin type II, serin proteinase inhibitor and transthyretin. After correction of spot volumes by total protein concentrations, 10 spots derived from albumin, cytokeratin type II and alpha1-antitrypsin remained significantly increased.Conclusion: The proteomic composition of AH differed significantly between patients and controls. The identified proteins suggest that the changes in AH are due to at least three different mechanisms: breakdown of the aqueous-blood barrier, enzymatic degradation, and liberation of locally synthesized proteins.