figure 3 (IMAGE)
Caption
(left) In a cell-free enzymatic reaction, 1-MNA is produced by NNMT in a time-dependent manner. P6A is used to monitor production of 1-MNA in this system. (right) P6A is also useful to detect 1- MNA in a crude biological sample. In animals, the concentration of urinary 1-MNA is highly dependent on the activity of NNMT and the total uptake of nicotinamide from foods. Therefore, intake of nicotinamide leads to an increase in the urinary excretion of 1- MNA in wild-type mice, while no 1-MNA excretion is detected in the Nnmt deficient mice. The fluorescence intensity of P6A is significantly quenched by wild-type derived urine samples. In addition, the fluorescence intensity was quenched by 1-MNA in a dose-dependent manner. (LC-MS/MS: quantification data using LC-MS/MS, P6A: data from fluorescence spectra with P6A, Wt: wild-type mice, Wt + NA: nicotinamide treatment group, Nnmt KO: NNMT deficient mice)
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Kanazawa University
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