Overexpression of the ABC Transporter BmrA Within Intracellular Caveolae in Escherichia coli

Alexia Gobet (1) , Christine Jaxel (2) , Sandrine Magnard (1) , Manuel Garrigos (2) , Stéphane Orlowski (2) , Nadège Jamin (2) , Pierre Falson (1) , Vincent Chaptal (1)

1 MMSB - Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry
2 I2BC - Institut de Biologie Intégrative de la Cellule

We described here the overproduction and oriented membrane insertion of membrane protein inside intracellular vesicles named heterologous caveolae within E. coli. The method is described with BmrA, a multidrug efflux pump from Bacillus subtilis. BmrA is produced in these vesicles, thanks to the co-expression with the canine caveolin-1b, one of the two isoforms of caveolin-1. Enriched by sucrose gradient, the caveolae-containing fraction allows to probe the ATPase and Hoechst 33342 transport activities, the latter displaying a higher specific activity than the same without caveolin-1b.

Mots clés: ABC transporter membrane protein overexpression membrane caveolae cytoplasmic vesicles; ABC transporter; membrane protein overexpression; membrane caveolae; cytoplasmic vesicles

Domaines: Sciences du Vivant [q-bio]

Alexia Gobet, Christine Jaxel, Sandrine Magnard, Manuel Garrigos, Stéphane Orlowski, et al.. Overexpression of the ABC Transporter BmrA Within Intracellular Caveolae in Escherichia coli. Heterologous Expression of Membrane Proteins, 2507, Springer US, pp.79-89, 2022, Methods in Molecular Biology, ⟨10.1007/978-1-0716-2368-8_5⟩. ⟨hal-04112236⟩