Mechanism of Hsp104/ClpB inhibition by prion curing Guanidinium hydrochloride

► GdnHCl inhibits Hsp104 and ClpB via the same mechanism. ► GdnHCl binding to Hsp104/ClpB abrogates Hsp70 cooperation. ► GdnHCl inhibits continuous but not single ATP turnover at AAA-1.