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This lecture by the instructor delves into the molecular mechanisms of protein regulation within the Hsp90 chaperone machinery. Starting with a discussion on equilibrium versus steady state, the lecture covers the role of Hsp90 in signal transduction pathways, the use of single molecule FRET experiments, and the effects of inhibitors on Hsp90's open-close equilibrium. The lecture also explores the impact of drugs and inhibitors on cochaperone and substrate binding, as well as the concept of cooperativity in the ATPases of Hsp90. Through detailed analyses and experiments, the lecture provides insights into the complex regulatory mechanisms governing protein function at various scales.