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Methods Mol Biol. 2017;1615:199-210. doi: 10.1007/978-1-4939-7033-9_16.

Fusion Reporter Approaches to Monitoring Transmembrane Helix Interactions in Bacterial Membranes.

Author information

1
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ-CNRS, 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France.
2
Division of Infectious Diseases and Harvard Medical School, Department of Microbiology and Immunobiology, Howard Hughes Medical Institute, Brigham and Women's Hospital, Boston, MA, USA.
3
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ-CNRS, 31 Chemin Joseph Aiguier, 13402, Marseille Cedex 20, France. cascales@imm.cnrs.fr.

Abstract

In transenvelope multiprotein machines such as bacterial secretion systems, protein-protein interactions not only occur between soluble domains but might also be mediated by helix-helix contacts in the inner membrane. Here we describe genetic assays commonly used to test interactions between transmembrane α-helices in their native membrane environment. These assays are based on the reconstitution of dimeric regulators allowing the control of expression of reporter genes. We provide detailed protocols for the TOXCAT and GALLEX assays used to monitor homotypic and heterotypic transmembrane helix-helix interactions.

KEYWORDS:

GALLEX; Helix–helix interaction; Membrane protein; One-hybrid; Protein-protein interaction; TOXCAT; Transmembrane segment; Two-hybrid; cI repressor

PMID:
28667614
DOI:
10.1007/978-1-4939-7033-9_16
[Indexed for MEDLINE]

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