Analyzing Persister Proteomes with SILAC and Label-Free Methods

Abstract

State-of-the-art mass spectrometry enables in-depth analysis of proteomes in virtually all organisms. This chapter describes methods for the analysis of persister proteomes by mass spectrometry. Stable isotope labeling by amino acids in cell culture (SILAC) is applied to assess protein biosynthesis in persister cells, which are isolated by treatment with beta-lactam antibiotics. Furthermore, persister proteomes during the postantibiotic recovery phase are analyzed by label-free quantification. The presented methods are valuable tools to shed light on persister physiology.

Keywords: Label-free quantification; Mass spectrometry; Persister cells; Postantibiotic recovery; Proteomics; SILAC.