Together with the Hsp70 system, in a regulated, concerted manner, the Hsp90 machinery processes a variety of different client proteins in a plethora of pathways, herein ensuring proteostasis. In this work, using different biophysical and biochemical assays, the interaction of the full-length glucocorticoid receptor with the chaperone systems via distinct conformational rearrangements was investigated. Moreover, the conformational dynamics of the Hsp90 chaperone regarding its catalytic activity and the methylation of a specific lysine in the C-terminal domain were studied.