Chapitre D'ouvrage Année : 2017

Recombinant Overexpression of Mammalian TSPO Isoforms 1 and 2

Résumé

TSPO is a 18 kDa membrane protein that exists in mammalian as two isoforms 1 and 2. They are involved in different functions and are located in different membranes. TSPO1 is mainly located in outer mitochondrial membrane, whereas TSPO2 is encountered in plasma membrane of red blood cells. Determination of their structures is a milestone to understand their function. Their natural abundance is not sufficient to get large amounts usually required for structural studies. We described heterologous overexpression in both bacterial and cell-free system and purification on immobilized-metal affinity chromatography (IMAC) of both proteins. Using the same vector, TSPO1 is mostly recovered in bacterial inclusion bodies whereas TSPO2 is found in both bacterial cytosol and inclusion bodies, but in low amounts. Cell-free expression was the best system to overexpress pure TSPO2.

Fichier non déposé

Dates et versions

hal-05452323 , version 1 (10-01-2026)

Identifiants

Citer

Lucile Senicourt, Soria Iatmanen-Harbi, Claude Hattab, Mariano Anibal Ostuni, Marie-France Giraud, et al.. Recombinant Overexpression of Mammalian TSPO Isoforms 1 and 2. Membrane Protein Structure and Function Characterization, 1635, Springer New York, pp.1-25, 2017, Methods in Molecular Biology, ⟨10.1007/978-1-4939-7151-0_1⟩. ⟨hal-05452323⟩
18 Consultations
2 Téléchargements

Altmetric

Partager

  • More