FIGURE 4.

GTN-triggered activation of purified sGC in the presence of vascular smooth muscle cell lysates. 150 μg of cell lysates from VSMC expressing either WT or C301S/C303S ALDH2 were incubated at 37 °C for 10 min in the presence of 50 ng of sGC, [α-³²P]GTP (∼200,000 cpm), 1,000 units/ml SOD, 1 mm NAD⁺, 2 mm DTT, and the indicated concentrations of GTN (10 nm to 100 μm), followed by isolation and quantification of cGMP as described under “Experimental Procedures.” Data were normalized to ALDH2 content to account for different expression levels and represent mean values ± S.E. of 3–4 independent experiments (*, p < 0.05).