Fiche publication
Date publication
janvier 2018
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr RUFF Marc
Tous les auteurs :
Eiler S, Levy N, Maillot B, Batisse J, Aubreton KP, Oladosu O, Ruff M
Lien Pubmed
Résumé
Purification of proteins containing disordered regions and participating in transient complexes is often challenging because of the small amounts available after purification, their heterogeneity, instability, and/or poor solubility. To circumvent these difficulties, we set up a methodology that enables the production of stable complexes in large amounts for structural and functional studies. In this chapter, we describe the methodology used to establish the best cell culture conditions and buffer compositions to optimize soluble protein production and their stabilization through protein complex formation. Two examples of challenging protein families are described, namely, the human steroid nuclear receptors and the HIV-1 pre-integration complexes.
Mots clés
GR, HIV, Integrase, Nuclear receptor, Pre-integration complex, Protein complex, TIF2
Référence
Methods Mol. Biol.. 2018 ;1764:315-328
