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二维凝胶电泳分析人正常尿液的可变性揭示了“公共”和“私人”蛋白质组。

Analysis of the variability of human normal urine by 2D-GE reveals a "public" and a "private" proteome.

机构信息

Sysdiag UMR 3145 CNRS Bio-Rad, Cap delta/Parc euromédecine, 1682 rue de la Valsière CS 61003, Montpellier, France.

出版信息

J Proteomics. 2011 Dec 10;75(1):70-80. doi: 10.1016/j.jprot.2011.06.031. Epub 2011 Jul 20.

Abstract

The characterization of the normal urinary proteome is steadily progressing and represents a major interest in the assessment of clinical urinary biomarkers. To estimate quantitatively the variability of the normal urinary proteome, urines of 20 healthy people were collected. We first evaluated the impact of the sample conservation temperature on urine proteome integrity. Keeping the urine sample at RT or at +4°C until storage at -80°C seems the best way for long-term storage of samples for 2D-GE analysis. The quantitative variability of the normal urinary proteome was estimated on the 20 urines mapped by 2D-GE. The occurrence of the 910 identified spots was analysed throughout the gels and represented in a virtual 2D gel. Sixteen percent of the spots were found to occur in all samples and 23% occurred in at least 90% of urines. About 13% of the protein spots were present only in 10% or less of the samples, thus representing the most variable part of the normal urinary proteome. Twenty proteins corresponding to a fraction of the fully conserved spots were identified by mass spectrometry. In conclusion, a "public" urinary proteome, common to healthy individuals, seems to coexist with a "private" urinary proteome, which is more specific to each individual.

摘要

正常尿液蛋白质组的特征分析正在不断深入,这对临床尿液生物标志物的评估具有重要意义。为了定量估计正常尿液蛋白质组的可变性,收集了 20 名健康人的尿液。我们首先评估了样本保存温度对尿液蛋白质组完整性的影响。将尿液样本保持在 RT 或+4°C 直到 -80°C 保存,对于 2D-GE 分析的长期储存样本似乎是最好的方法。通过 2D-GE 对 20 个尿液样本进行了映射,对正常尿液蛋白质组的定量可变性进行了估计。通过分析整个凝胶中的 910 个鉴定斑点,并将其表示在虚拟 2D 凝胶中,分析了其出现情况。发现 16%的斑点出现在所有样本中,23%出现在至少 90%的尿液中。约 13%的蛋白质斑点仅出现在 10%或更少的样本中,因此代表了正常尿液蛋白质组中最易变的部分。通过质谱鉴定出对应于完全保守斑点的一部分的 20 种蛋白质。总之,健康个体共有的“公共”尿液蛋白质组似乎与每个个体更特异的“私人”尿液蛋白质组共存。

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