Fiche publication
Date publication
février 2008
Auteurs
Membres identifiés du Cancéropôle Est :
Pr WEISS Etienne
,
Pr DIDIER Pascal
Tous les auteurs :
Didier P, Weiss E, Sibler AP, Philibert P, Martineau P, Bigot JY, Guidoni L
Lien Pubmed
Résumé
Time-resolved femtosecond spectroscopy can improve the application of green fluorescent proteins (GFPs) as protein-folding reporters. The study of ultrafast excited-state dynamics (ESD) of GFP fused to single chain variable fragment (scFv) antibody fragments, allowed us to define and measure an empirical parameter that only depends on the folding quality (FQ) of the fusion. This method has been applied to the analysis of genetic fusions expressed in the bacterial cytoplasm and allowed us to distinguish folded and thus functional antibody fragments (high FQ) with respect to misfolded antibody fragments. Moreover, these findings were strongly correlated to the behavior of the same scFvs expressed in animal cells. This method is based on the sensitivity of the ESD to the modifications in the tertiary structure of the GFP induced by the aggregation state of the fusion partner. This approach may be applicable to the study of the FQ of polypeptides over-expressed under reducing conditions.
Référence
Biochem Biophys Res Commun. 2008 Feb 22;366(4):878-84
