(4) Compound X is an inhibitor of mitochondrial ATP synthesis. It was observed that when compound X was added to cells, the NAD*/NADH ratio decreased. Would you expect X to be an uncoupling agent or an inhibitor of the respiratory chain? Please explain. (4%)

1. A scientist uses the yeast expression system to over-express one recombinant protein, and which is also an enzyme. Based on the enzymatic activity of the recombinant protein, the scientist is able to purify the protein and makes the purification table. Purification procedure Total protein (µg) Enzymatic activity (units) 1. Crude extract 60,000 1,000,000 2. Sodium sulfate precipitation 30,000 700,000 3. Affinity chromatography 5,000 520,000 4. Ion-exchange chromatography 3,800 376,000 (1) From the information given in the table, please calculate the specific activity of the enzyme after the final procedure. (2%)

(4) Compound X is an inhibitor of mitochondrial ATP synthesis. It was observed that when compound X was added to cells, the
NAD*/NADH ratio decreased. Would you expect X to be an uncoupling agent or an inhibitor of the respiratory chain? Please
explain. (4%)

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