FIGURE 5.
Factors that are required for heme insertion into apo-sGC also drive the NO effect on hsp90 binding. A, B, and D, Western analyses of immunoprecipitations of sGC-β1 showing how 50 μm SNAP treatment for indicated times impacts level of hsp90 associated with apo-sGC-β1 expressed in heme-deficient (+SA) RFL-6 or COS-7 cells or with the sGC-β1H105F heme-free mutant expressed in heme-replete COS-7 cells (input 20%). C, cGMP levels in supernatants of cells in A and B. E and F, impact of an hsp90 inhibitor (radicicol; Rad) on hsp90-sGC-β1 association levels and cGMP production in cells given a 5-min SNAP treatment. Values are mean ± S.D. of three independent experiments (*, p < 0.05, by one-way ANOVA). IB, immunoblot.