One-step affinity purification of fusion proteins with optimal monodispersity and biological activity: application to aggregation-prone HPV E6 proteins.
Fiche publication
Date publication
décembre 2018
Journal
Microbial cell factories
Auteurs
Membres identifiés du Cancéropôle Est :
Dr TRAVE Gilles, Dr NOMINE Yves
Tous les auteurs :
Bonhoure A, Demenge A, Kostmann C, San José L, De la Cal E, Armisen P, Nominé Y, Travé G
Lien Pubmed
Résumé
Bacterial expression and purification of recombinant proteins under homogeneous active form is often challenging. Fusion to highly soluble carrier proteins such as Maltose Binding Protein (MBP) often improves their folding and solubility, but self-association may still occur. For instance, HPV E6 oncoproteins, when produced as MBP-E6 fusions, are expressed as mixtures of biologically inactive oligomers and active monomers. While a protocol was previously developed to isolate MBP-E6 monomers for structural studies, it allows the purification of only one MBP-E6 construct at the time. Here, we explored a parallelizable strategy more adapted for biophysical assays aiming at comparing different E6 proteins.
Mots clés
IMAC, MBP fusion, One-step purification, Protein aggregation, Protein solubility
Référence
Microb. Cell Fact.. 2018 Dec 1;17(1):191