Active unfolding of the glucocorticoid receptor by the Hsp70/Hsp40 chaperone system in single-molecule mechanical experiments

Abstract

Significance One of the key unresolved questions in the field of molecular chaperones is how they can actively unfold proteins. In this study, we discovered that the Hsp70/Hsp40 chaperone system completely unfolds a native soluble substrate protein, the ligand-binding domain of the glucocorticoid receptor, in a concerted action. Our high-resolution optical tweezers data show in real time how the substrate is attacked by the chaperone machinery. As soon as the hormone has left the binding pocket, up to five Hsp70/Hsp40 complexes bind and unfold the protein in a stepwise manner. This finding constitutes direct evidence that the chaperone machinery can bind to the folded core of the receptor, thus providing a mechanism for Hsp70-induced protein unfolding.