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正常人类尿液足细胞外泌体富集部分的蛋白质组学分析。

Proteomic analysis of podocyte exosome-enriched fraction from normal human urine.

机构信息

Institute of Clinical Pathology, Geneva University Hospitals, Geneva CH-1211, Switzerland.

出版信息

J Proteomics. 2013 Apr 26;82:193-229. doi: 10.1016/j.jprot.2013.01.012. Epub 2013 Jan 30.

Abstract

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.

摘要

尿液来源于肾脏肾小球和肾小管部分的协同活动。作为这些细胞功能过程的足迹,尿外泌体是肾脏上皮细胞通过与质膜融合并释放到细胞外环境中的 40-80nm 膜囊泡,是鉴定蛋白质并研究其在肾脏中作用的来源。本研究的目的是基于用足细胞特异性 CR1 免疫包被珠进行尿免疫吸附,用 LC-MS/MS 技术进行蛋白质组学分析,鉴定足细胞外泌体蛋白。该方法鉴定出 1195 种蛋白质。通过生物信息学方法,鉴定出 27 种脑表达蛋白,其中 14 种蛋白在肾脏中以 mRNA 水平表达,其中一种蛋白 COMT 蛋白在足细胞中以蛋白水平表达。这些结果证明了该方法鉴定足细胞蛋白的可靠性,现在需要通过进一步的实验来更精确地分析它们在足细胞中的生物学功能。

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