TY - JOUR T1 - Interaction between the 90-kDa Heat Shock Protein and Soluble Guanylyl Cyclase: Physiological Significance and Mapping of the Domains Mediating Binding<sup/> JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1133 LP - 1141 DO - 10.1124/mol.105.012682 VL - 68 IS - 4 AU - Andreas Papapetropoulos AU - Zongmin Zhou AU - Christina Gerassimou AU - Gunay Yetik AU - Richard C. Venema AU - Charis Roussos AU - William C. Sessa AU - John D. Catravas Y1 - 2005/10/01 UR - http://molpharm.aspetjournals.org/content/68/4/1133.abstract N2 - The 90-kDa heat shock protein (hsp90) regulates the stability and function of many client proteins, including members of the NO-cGMP signaling pathway. Soluble guanylyl cyclase (sGC), an NO receptor, was recently reported to be an hsp90-interacting partner. In the present study, we show that hsp90 binds to both subunits of the most common sGC form (α1β1) when these are expressed individually but only interacts with β1 in the heterodimeric form of the enzyme. Characterization of the region of hsp90 required to bind each subunit in immunoprecipitation experiments revealed that residues 310 to 456 of hsp90 interact with the sGC subunits. The region of β1 responsible for binding to hsp90β was mapped using in vitro binding assays and immunoprecipitation experiments and was found to lie in the regulatory domain. The physiological importance of the hsp90/sGC interaction was investigated by treating rat smooth muscle cells with the hsp90 inhibitors radicicol and geldanamycin (GA) and determining both sGC activity and protein levels. Long-term (24 or 48 h) inhibition of hsp90 resulted in a strong decrease of both α1 and β1 protein levels and sGC activity. Moreover, incubation of smooth muscle cells with the proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132) blocked the GA-induced down-regulation of sGC. We conclude that the N-terminal region of the β1 subunit mediates binding of the heterodimeric form of sGC to hsp90 and that this interaction involves the M domain of hsp90. Hsp90 binding to sGC regulates the pool of active enzymes by affecting the protein levels of the two subunits. ER -