Proteomic analysis of podocyte exosome-enriched fraction from normal human urine
Proteomic analysis of podocyte exosome-enriched fraction from normal human urine
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DOI：
10.1016/j.jprot.2013.01.012
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发表时间：
2013-04-26
影响因子：
3.3
通讯作者：
Moll, Solange
中科院分区：
文献类型：
--
作者：
Prunotto, Marco;Farina, Annarita;Moll, Solange
尿液是肾脏肾小球和肾小管室协调活动的结果。作为这些细胞功能过程的足迹，尿外泌体以及肾上皮细胞与质膜融合后释放到细胞外环境中的 40-80 nm 膜囊泡，是蛋白质鉴定和研究其在肾脏中的作用的来源。本研究的目的是使用足细胞特异性 CR1 免疫涂层珠子基于尿液免疫吸收来鉴定足细胞外泌体蛋白，然后使用 LC MS/MS 技术进行蛋白质组分析。该方法可鉴定 1195 种蛋白质。通过生物信息学方法，鉴定了27种脑表达蛋白，其中14种新被证明在肾脏中以mRNA水平表达，其中一种COMT蛋白被证明在足细胞中以蛋白质水平表达。这些结果证明了识别足细胞蛋白的方法的可靠性，现在需要通过进一步的实验来完成，以更精确地分析它们在足细胞中的生物学功能。 (C) 2013 Elsevier B.V. 保留所有权利。
Urine results from a coordinated activity of glomerular and tubular compartments of the kidney. As a footprint of these cellular functional processes, urinary exosomes, and 40-80 nm membrane vesicles released after fusion with the plasma membrane into the extracellular environment by renal epithelial cells, are a source for identification of proteins and investigation of their role in the kidney. The aim of the present study was the identification of podocyte exosome proteins based on urine immunoabsorption using podocyte-specific CR1-immunocoated beads followed by proteomic analysis using LC MS/MS techniques. This methodology allowed the identification of 1195 proteins. By using a bioinformatic approach, 27 brain-expressed proteins were identified, in which 14 out of them were newly demonstrated to be expressed in the kidney at a mRNA level, and, one of them, the COMT protein, was demonstrated to be expressed in podocytes at a protein level. These results, attesting the reliability of the methodology to identify podocyte proteins, need now to be completed by further experiments to analyze more precisely their biological function(s) in the podocytes. (C) 2013 Elsevier B.V. All rights reserved.