Figure 8. Role of the 669WTAPELL675 motif in ANF-RGC signal transduction.
(A) Western analysis. COS cells were transfected with wt ANF-RGC or its 669WTAPELL675 deletion mutant and their particulate fractions were prepared as described in ”Materials and Methods”. The membranes were analyzed by Western blotting using antibodies against ANF-RGC. Lane1, wt ANF-RGC; Lane 2, 669WTAPELL675 deletion mutant. (B) ANF/ATP-dependent cyclase activity of ANF-RGC. Wild type ANF-RGC and the deletion mutant were individually expressed in COS cells and their membranes were analyzed for ANF/ATP-dependent cyclase activity. Cyclic GMP formed was measured by radioimmunoassay. The experiment was done in triplicate and repeated two times for reproducibility. The results presented (mean ± SD) are from these experiments.