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用于分析人肾结石基质成分的无标记蛋白质组学方法。

Label-free proteomic methodology for the analysis of human kidney stone matrix composition.

作者信息

Witzmann Frank A, Evan Andrew P, Coe Fredric L, Worcester Elaine M, Lingeman James E, Williams James C

机构信息

Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 635 Barnhill Drive, Room 362A, Indianapolis, IN 46202-5120 USA.

Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN USA.

出版信息

Proteome Sci. 2016 Feb 27;14:4. doi: 10.1186/s12953-016-0093-x. eCollection 2016.

DOI:10.1186/s12953-016-0093-x
PMID:26924944
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4769560/
Abstract

BACKGROUND

Kidney stone matrix protein composition is an important yet poorly understood aspect of nephrolithiasis. We hypothesized that this proteome is considerably more complex than previous reports have indicated and that comprehensive proteomic profiling of the kidney stone matrix may demonstrate relevant constitutive differences between stones. We have analyzed the matrices of two unique human calcium oxalate stones (CaOx-Ia and CaOx-Id) using a simple but effective chaotropic reducing solution for extraction/solubilization combined with label-free quantitative mass spectrometry to generate a comprehensive profile of their proteomes, including physicochemical and bioinformatic analysis.`.

RESULTS

We identified and quantified 1,059 unique protein database entries in the two human kidney stone samples, revealing a more complex proteome than previously reported. Protein composition reflects a common range of proteins related to immune response, inflammation, injury, and tissue repair, along with a more diverse set of proteins unique to each stone.

CONCLUSION

The use of a simple chaotropic reducing solution and moderate sonication for extraction and solubilization of kidney stone powders combined with label-free quantitative mass spectrometry has yielded the most comprehensive list to date of the proteins that constitute the human kidney stone proteome.

摘要

背景

肾结石基质蛋白组成是肾结石病一个重要但却了解甚少的方面。我们推测，这种蛋白质组比之前报道的要复杂得多，并且对肾结石基质进行全面的蛋白质组分析可能会揭示不同结石之间相关的组成差异。我们使用一种简单但有效的离液剂还原溶液进行提取/溶解，并结合无标记定量质谱分析，对两块独特的人草酸钙结石（CaOx-Ia和CaOx-Id）的基质进行了分析，以生成其蛋白质组的全面图谱，包括物理化学和生物信息学分析。

结果

我们在两个人肾结石样本中鉴定并定量了1059个独特的蛋白质数据库条目，揭示了一个比之前报道更为复杂的蛋白质组。蛋白质组成反映了一系列与免疫反应、炎症、损伤和组织修复相关的常见蛋白质，以及每块结石特有的更多样化的蛋白质。

结论

使用简单的离液剂还原溶液和适度超声处理来提取和溶解肾结石粉末，并结合无标记定量质谱分析，得到了迄今为止构成人肾结石蛋白质组的最全面的蛋白质列表。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9051/4769560/0aaa7d356ef6/12953_2016_93_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9051/4769560/71e698f8b84e/12953_2016_93_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9051/4769560/0aaa7d356ef6/12953_2016_93_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9051/4769560/71e698f8b84e/12953_2016_93_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9051/4769560/0aaa7d356ef6/12953_2016_93_Fig2_HTML.jpg

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