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Triflic Acid Treatment Enables LC-MS/MS Analysis of Insoluble Bacterial Biomass.

ABSTRACT: 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.

SUBMITTER: Wang AY

PROVIDER: S-EPMC7108958 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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Triflic Acid Treatment Enables LC-MS/MS Analysis of Insoluble Bacterial Biomass.

Wang Ana Y AY Thuy-Boun Peter S PS Stupp Gregory S GS Su Andrew I AI Wolan Dennis W DW

Journal of proteome research 20180808 9

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 ...[more]

PMID: 30019906

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