Fusion Reporter Approaches to Monitoring Transmembrane Helix Interactions in Bacterial Membranes

Part of the Methods in Molecular Biology book series (MIMB, volume 1615)

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.

Key words

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

Notes

Acknowledgements

Work in the EC laboratory is supported by the Centre National de la Recherche Scientifique, the Aix-Marseille Université, and grants from the Agence Nationale de la Recherche (ANR-14-CE14-0006-02 and ANR-15-CE11-0019-01). LL and AZ are recipients of doctoral fellowships from the French Ministère de l’Enseignement Supérieur et de la Recherche and end-of-thesis fellowships from the Fondation pour la Recherche Médicale (FDT20160435498 and FDT20140931060).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Laureen Logger
    • 1
  • Abdelrahim Zoued
    • 1
    • 2
  • Eric Cascales
    • 1
  1. 1.Laboratoire d’Ingénierie des Systèmes Macromoléculaires, UMR7255Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ—CNRSMarseille Cedex 20France
  2. 2.Division of Infectious Diseases and Harvard Medical School, Department of Microbiology and ImmunobiologyHoward Hughes Medical Institute, Brigham and Women’s HospitalBostonUSA

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