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Triflic Acid Treatment Enables LC-MS/MS Analysis of Insoluble Bacterial Biomass.
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2018-08-08 , DOI: 10.1021/acs.jproteome.8b00166
Ana Y Wang 1 , Peter S Thuy-Boun 1 , Gregory S Stupp 1 , Andrew I Su 1 , Dennis W Wolan 1
Affiliation  

The lysis and extraction of soluble bacterial proteins from cells is a common practice for proteomics analyses, but insoluble bacterial biomasses are often left behind. Here, we show that with triflic acid treatment, the insoluble bacterial biomass of Gram- and Gram+ bacteria can be rendered soluble. We use LC-MS/MS shotgun proteomics to show that bacterial proteins in the soluble and insoluble postlysis fractions differ significantly. Additionally, in the case of Gram- Pseudomonas aeruginosa, triflic acid treatment enables the enrichment of cell-envelope-associated proteins. Finally, we apply triflic acid to a human microbiome sample to show that this treatment is robust and enables the identification of a new, complementary subset of proteins from a complex microbial mixture.

中文翻译:

三氟甲磺酸处理可以对不溶性细菌生物质进行LC-MS / MS分析。

从细胞中裂解和提取可溶性细菌蛋白是蛋白质组学分析的常用方法,但不溶性细菌生物量通常会留下。在这里,我们表明,通过三氟甲磺酸处理,可以使革兰氏和革兰氏阳性细菌的不溶细菌生物量变得可溶。我们使用LC-MS / MS shot弹枪蛋白质组学来显示可溶和不可溶的后分解组分中的细菌蛋白存在显着差异。另外,在绿脓杆菌中,三氟甲磺酸处理可以富集细胞包膜相关蛋白。最后,我们将三氟甲磺酸应用于人的微生物组样品,以表明这种处理是可靠的,并且能够从复杂的微生物混合物中鉴定出新的互补蛋白质子集。
更新日期:2018-08-08
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