Research Article

A Regulatory Role of NAD Redox Status on Flavin Cofactor Homeostasis in S. cerevisiae Mitochondria

Figure 2

Fluorimetric evidence of NADH cleavage catalyzed by solubilized SCM. (a) SCM (0.025 mg protein) were added to NADH (5 μM) in the standard medium supplemented with KCN (1 mM) and the reaction followed at 25°C, essentially as described in Section 2. NADH fluorescence excitation spectra (emission wavelength set at 456 nm) were recorded at different incubation times. (b) NADH fluorescence decrease, induced by freezing-thawing rupture of SCM, was continuously measured at both the wavelength pairs 260/456 and 340/456 nm, under the experimental condition described in (a). Where indicated NaF or CaCl2 (1 mM each) were added (dotted lines).
612784.fig.002a
(a)
612784.fig.002b
(b)