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Measurement of FMN, riboflavin, or NADH binding by ITC experiments. The trend of heat released by serial injections of FMN (▪), riboflavin (▾), or NADH (•) into EmoB was monitored. FMN showed the typical heat-releasing pattern. Neither riboflavin nor NADH had any detectable heat-releasing events upon injections. Solid lines represent the least-square fits of the data using a single-site binding model.
