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J Proteome Res. 2018 Sep 7;17(9):2978-2986. doi: 10.1021/acs.jproteome.8b00166. Epub 2018 Aug 8.

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

Author information

1
Department of Molecular Medicine and Department of Integrative Structural and Computational Biology , The Scripps Research Institute , 10550 North Torrey Pines Road , La Jolla , California 92037 , United States.

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.

KEYWORDS:

ComPIL; MudPIT; membrane proteins; metaproteomics; microbiome; triflic acid

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