Table 2.

Binding free energies and equilibrium dissociation constant of Dbh and the mutants to primer/template DNA.

Mutation sites	Primer/Template DNA(a)	Free binding energy Δ Gbind (kcal/mol)	(ΔGbind)mutant − (ΔGbind)wt (kcal/mol)	Variants	Equilibrium dissociation constant Kd(nM)
WTDbh	P	−2251.8	0	WTDbh	54.8 ± 3.8
	T	−3465.7		Sdbh	51.2 ± 2.4
KSKIP(241–245) RVRKS	P	−2579.5	−532.2	SdbhKSKIP(241–245) RVRKS	34.8 ± 1.2
	T	−3670.2
M76I	P	−2414.7	−380	SdbhM76I	42.8 ± 3.1
	T	−3682.8
L250V	P	−2422.1	−318.3	SdbhL250V	50.4 ± 3.4
	T	−3603.7
T37F	P	−2313.6	−159	SdbhT37F	49.7 ± 3.1
	T	−3562.9
A221S	P	−2344.03	−100.45	SdbhA221S	47.5 ± 3.0
	T	−3473.91
I62V	P	−2145.6	−60.3	SdbhI62V	49.8 ± 2.6
	T	−3632.2
K337R	P	−2554.6	42.4	SdbhK337R	55.5 ± 4.1
	T	−3120.5
Y249I	P	−2305.0	293.6	SdbhY249I	56.2 ± 3.4
	T	−3118.9

^(a)dsDNA (Primer/Template DNA) referred to the chain primer extended (P(5-3′)) and the corresponding template chain (T(3′-5′)).