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大气气溶胶样品的代谢组学分析。

Metaproteomic analysis of atmospheric aerosol samples.

机构信息

Department of Multiphase Chemistry, Max Planck Institute for Chemistry, Hahn-Meitner-Weg 1, 55128, Mainz, Germany.

School of Environment and Energy, South China University of Technology, Higher Education Mega Center, Guangzhou, 510006, People's Republic of China.

出版信息

Anal Bioanal Chem. 2016 Sep;408(23):6337-48. doi: 10.1007/s00216-016-9747-x. Epub 2016 Jul 13.

Abstract

Metaproteomic analysis of air particulate matter provides information about the abundance and properties of bioaerosols in the atmosphere and their influence on climate and public health. We developed and applied efficient methods for the extraction and analysis of proteins from glass fiber filter samples of total, coarse, and fine particulate matter. Size exclusion chromatography was applied to remove matrix components, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied for protein fractionation according to molecular size, followed by in-gel digestion and LC-MS/MS analysis of peptides using a hybrid Quadrupole-Orbitrap MS. Maxquant software and the Swiss-Prot database were used for protein identification. In samples collected at a suburban location in central Europe, we found proteins that originated mainly from plants, fungi, and bacteria, which constitute a major fraction of primary biological aerosol particles (PBAP) in the atmosphere. Allergenic proteins were found in coarse and fine particle samples, and indications for atmospheric degradation of proteins were observed. Graphical abstract Workflow for the metaproteomic analysis of atmospheric aerosol samples.

摘要

大气颗粒物的宏蛋白质组学分析提供了有关大气中生物气溶胶的丰度和特性及其对气候和公众健康影响的信息。我们开发并应用了从总悬浮颗粒物、粗颗粒物和细颗粒物的玻璃纤维滤膜样品中提取和分析蛋白质的有效方法。采用排阻色谱法去除基质成分,然后根据分子大小进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)进行蛋白质分级,接着使用混合四极杆轨道阱质谱对肽进行胶内消化和 LC-MS/MS 分析。使用 Maxquant 软件和 Swiss-Prot 数据库进行蛋白质鉴定。在中欧郊区位置采集的样品中,我们发现了主要来源于植物、真菌和细菌的蛋白质,这些蛋白质构成了大气中初级生物气溶胶颗粒(PBAP)的主要部分。在粗颗粒和细颗粒样品中发现了过敏原蛋白,并观察到了蛋白质在大气中降解的迹象。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8271/5009178/c6b21ec255ae/216_2016_9747_Figa_HTML.jpg

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