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Methods Mol Biol. 2017;1615:459-464. doi: 10.1007/978-1-4939-7033-9_31.

Identification of Effectors: Precipitation of Supernatant Material.

Author information

1
Laboratoire d'Ingénierie des Systèmes Macromoléculaires (LISM, UMR7255), Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France.
2
Laboratoire d'Ingénierie des Systèmes Macromoléculaires (LISM, UMR7255), Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France. ljournet@imm.cnrs.fr.

Abstract

Bacterial secretion systems allow the transport of proteins, called effectors, as well as external machine components in the extracellular medium or directly into target cells. Comparison of the secretome, i.e. the proteins released in the culture medium, of wild-type and mutant cells provides information on the secretion profile. In addition, mass spectrometry analyses of the culture supernatant of bacteria grown in liquid culture under secreting conditions allows the identification of secretion system substrates. Upon identification of the substrates, the secretion profile serves as a tool to test the functionality of secretion systems. Here we present a classical method used to concentrate the culture supernatant, based on trichloroacetic acid precipitation.

KEYWORDS:

Secretome; Supernatant; TCA precipitation

PMID:
28667629
DOI:
10.1007/978-1-4939-7033-9_31
[Indexed for MEDLINE]

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