Figure 2. Citrulline formation as a function of CaM concentrations.

Citrulline forming activity was determined in a reaction mixture containing 25 mM Tris, pH 7.5, 100 nM eNOS, 3H-L-arginine (1μCi), L-arginine (20 μM), NADPH (100μM), H₄B (10μM), and CaCl₂ (300 μM), 0.2 mM EDTA, 100 mM NaCl, 0.1 mM DTT, 10% glycerol and the indicated concentration of each CaM construct as described under Experimental Procedures. Data are mean of duplicate measurements with <5% variation. Each experiment was repeated three times.