Table 2. Thermodynamic parameters determined for the interaction of embonic acid with human m-NAD(P)-ME.

Characteristics	m-NAD(P)-ME	aAverage N (sites)	bKd (μM)	cΔG (kcal/mol)	dΔH (kcal/mol)	fTΔS (kcal/mol)	eΔS (kcal/mol/deg)
	WT	3.8 ± 0.2	2.7 ± 0.7	−7.6	−4.6 ± 0.3	3.0	9.9
Tetramer interface mutant	H142A/D568A	1.4 ± 0.03	0.4 ± 0.1	−8.8	−12.3 ± 0.4	−3.5	−11.7
Exo site mutant (at tetramer interface)	R197E	3.4 ± 0.1	1.0 ± 0.2	−8.2	−5.1 ± 0.1	3.1	10.5
Dimer interface mutant	Q51A/E90A	1.8 ± 0.1	1.1 ± 0.4	−8.1	4.7 ± 0.4	12.8	42.8
Fumarate site mutant (at dimer interface)	E59N	2.0 ± 0.2	3.5 ± 1.1	−7.4	−1.7 ± 0.2	5.8	19.4
	R67A	1.9 ± 0.2	4.0 ± 0.7	−7.3	4.0 ± 0.2	11.4	38.1
	R91A	3.7 ± 0.4	13.1 ± 3.8	−6.7	−1.7 ± 0.3	5.0	16.5

^aAverage N (sites) signifies the stoichiometry of embonic acid in a tetrameric malic enzyme at equilibrium.

^bK_d (μM) represents the dissociation constant of embonic acid against m-NAD(P)-ME, which is reciprocal of K_A (association constant).

^cΔG (kcal/mol)

^dΔH (kcal/mol) and

^eΔS (kcal/mol/deg) represent the free energy change during the binding process.

^fT was fixed at 298K.