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85030890760
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Enzyme assays were performed as follows. A typical reaction (total volume 1.0 mL) contained 50 mM imidazole (pH=7.0), 1 mM EDTA, 0.05% (v/v) β-mercaptoethanol, 5% DMSO, 0.1-0.2 μM VH1 (or 0.5-1 μM VHR), and 0-2700 μM inhibitor. After incubation of the enzyme with the inhibitor for 1 h at room temperature, the reaction was initiated by the addition of 5 mM pNPP. The reaction progress was monitored at 405 nm on a Perkin-Elmer UV-Vis spectrophotometer
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Enzyme assays were performed as follows. A typical reaction (total volume 1.0 mL) contained 50 mM imidazole (pH=7.0), 1 mM EDTA, 0.05% (v/v) β-mercaptoethanol, 5% DMSO, 0.1-0.2 μM VH1 (or 0.5-1 μM VHR), and 0-2700 μM inhibitor. After incubation of the enzyme with the inhibitor for 1 h at room temperature, the reaction was initiated by the addition of 5 mM pNPP. The reaction progress was monitored at 405 nm on a Perkin-Elmer UV-Vis spectrophotometer.
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19
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85030905244
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note
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6 value was determined from the reactivation experiment as described in the text. All measurements were performed at room temperature.
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