Research Article

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

Figure 1

Purity, integrity, and functionality of SCM. (a) The amount of PDC, AP, D-AAOX, and CS activities in sphero and SCM (0.05–0.1 mg protein) were measured, as reported in Section 2. The values of the enzymatic activities are the mean (±SD) of three experiments performed with different cellular preparations. (b) FUM activity was measured in sphero, intact SCM (SCM − TX100), and solubilized SCM (SCM + TX100) (0.05 mg protein each) as reported in Section 2. The values are the mean (±SD) of three experiments performed with different cellular preparations. (c) A schematic representation of the electron transport along the respiratory chain starting from the external NADH dehydrogenases (Nde1p and Nde2p) is reported. Polarographic measurements of the NADH-dependent oxygen uptake rate in SCM (0.1 mg protein) were carried out as described in Section 2. The arrows indicate when the additions were made. The numbers along the trace refer to the oxygen uptake rate expressed as ngatoms O · min−1 · mg−1 mitochondrial protein.
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