Sample Preparation for Mass-Spectrometry Based Absolute Protein Quantification in Antibiotic Stress Research

Florian Bonn; Sandra Maass; Dörte Becher

doi:10.1007/978-1-4939-6634-9_17

Sample Preparation for Mass-Spectrometry Based Absolute Protein Quantification in Antibiotic Stress Research

Methods Mol Biol. 2017:1520:281-289. doi: 10.1007/978-1-4939-6634-9_17.

Authors

Florian Bonn¹, Sandra Maass¹, Dörte Becher²

Affiliations

¹ Department for Microbial Proteomics, Institute for Microbiology, Ernst-Moritz-Arndt University of Greifswald, Friedrich-Ludwig-Jahn-Straße 15, Greifswald, 17489, Germany.
² Department for Microbial Proteomics, Institute for Microbiology, Ernst-Moritz-Arndt University of Greifswald, Friedrich-Ludwig-Jahn-Straße 15, Greifswald, 17489, Germany. dbecher@uni-greifswald.de.

PMID: 27873259
DOI: 10.1007/978-1-4939-6634-9_17

Abstract

Absolute protein quantification is an essential tool for system biology approaches and elucidation of stoichiometry of multi-protein complexes. In this chapter, a universal protocol for gel free absolute protein quantification in bacterial systems is described, which can be used for sample preparation prior to miscellaneous mass-spectrometry-based quantification workflows like AQUA, Hi3, and emPAI. In addition, a focus has been set to the specific challenges in antibiotic stress research.

Keywords: AQUA; Absolute protein quantification; Antibiotic stress response; Gel free proteomics; Hi3; In-solution digestion; QconCAT; Sample preparation; emPAI.

MeSH terms

Anti-Bacterial Agents / pharmacology*
Bacterial Proteins / analysis*
Cell Fractionation
Colony Count, Microbial
Mass Spectrometry / methods*
Stress, Physiological*
Trypsin / metabolism

Substances

Anti-Bacterial Agents
Bacterial Proteins
Trypsin