Chapter 16. Fusion reporter approaches to monitor transmembrane helix interactions in bacterial membranes Running head: protein-protein interaction in the membrane

Laureen Logger (1) , Abdelrahim Zoued (1) , E. Cascales (1)

1 LISM - Laboratoire d'ingénierie des systèmes macromoléculaires

In transenvelope multiprotein machines such as bacterial secretion systems, protein-protein interactions not only occur between soluble domains but are also 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 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: membrane protein; protein-protein interaction; transmembrane segment; helix-; helix interaction; homotypic; heterotypic; one-hybrid; two-hybrid; cI repressor; TOXCAT; GALLEX; BACTH

Domains: Life Sciences [q-bio]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular biology; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Biochemistry [q-bio.BM]; Life Sciences [q-bio]; Microbiology and Parasitology; Bacteriology

Laureen Logger, Abdelrahim Zoued, E. Cascales. Chapter 16. Fusion reporter approaches to monitor transmembrane helix interactions in bacterial membranes Running head: protein-protein interaction in the membrane. Methods Mol Biol, 2017. ⟨hal-01780729⟩