Table 1. Effect of fumarate analogs on human m-NAD(P)-ME and c-NADP-ME^{1 , 2}.

	3 , 4m-NAD(P)-ME WT	3 , 4m-NAD(P)-ME R67A/R91A	3 , 4m-NAD(P)-ME K57S/E59N/K73E/D102S	3 , 5c-NADP-ME WT
Fumarate	1.98±0.14	0.98±0.06	0.97±0.04	1.01±0.04
Mesaconate	1.75±0.16	1.01±0.04	0.99±0.05	1.02±0.03
trans-Aconitate	1.15±0.03	1.01±0.05	0.98±0.03	0.99±0.04
Monomethyl fumarate	2.78±0.08	1.07±0.03	1.17±0.05	0.99±0.03
Monoethyl fumarate	1.83±0.04	0.98±0.04	1.01±0.05	1.01±0.03
Oxaloacetate	1.00±0.03	0.94±0.02	0.87±0.02	1.02±0.03
Diethyl oxalacetate	0.31±0.03	0.85±0.02	0.91±0.03	0.93±0.02
Dimethyl maleate	1.00±0.02	0.97±0.02	0.95±0.03	0.99±0.03
Diethyl maleate	0.88±0.03	0.99±0.03	0.95±0.03	1.01±0.02
Dimethyl fumarate	0.79±0.03	0.99±0.02	0.98±0.02	0.99±0.02
Diethyl fumarate	0.65±0.04	0.92±0.01	0.94±0.02	1.01±0.03

¹These values were the ratios of specific activities of the enzyme determined in the presence and absence of these chemical compounds.

²These values were the average with standard deviation of three-time repeats.

³The enzyme specific activities of the m-NADP-(P)-ME WT, R67A/R91A, K57S/E59N/K73E/D102S and c-NADP-ME WT were approximately 0.1, 0.005, 0.02 and 0.08 µmol/min, respectively, and the final enzyme concentration in an individual assay was 20, 400, 100, and 25 ng/µl, respectively.

⁴For m-NAD(P)-ME, the final concentrations of these analogs were fixed at 5 mM, except for monoethyl fumarate and mesaconate, which were fixed at 10 mM and 20 mM, respectively.

⁵For c-NADP-ME, the final concentrations of these analogs were fixed at 2 mM.