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. 2015 Nov 18;291(2):572–581. doi: 10.1074/jbc.M115.681221

FIGURE 5.

FIGURE 5.

AdoMet has no effect on CO binding to truncated CBS. A, CO titration profiles of reduced truncated CBS in the absence (solid symbols) or presence (open symbols) of 500 μm AdoMet. The two data sets were fitted to Equation 1 yielding Kd1 = 0.5 ± 0.4 μm (10%) and Kd2 = 16 ± 2 μm (90%), consistent with a C50 of 12 ± 1 μm. Experimental conditions are as in Fig. 3. B, reaction time courses acquired after stopped-flow mixing reduced truncated CBS (1.0 μm in heme) with 1 mm CO in the absence (dotted line) or presence of AdoMet (500 μm before mixing; solid line). Regardless of AdoMet, data fitting yielded the following rate constant (% reaction amplitude): k1 = 0.22 s−1 (20%) and k2 = 0.025 s−1 (80%). Inset, optical transitions obtained by global fit analysis of the spectral data acquired in the absence (dotted line) or presence of AdoMet (solid line).