Table 1. Thermodynamic parameters, microscopic rate constants and kinetic m^‡-values for monomeric variants.^*.

	WT	A4V	L38V	G93A	L106V	S134N
Apo reduceda
$\mathrm{\Delta }{\text{G}}_{(\text{U}/\text{M})}^{°}$ b	-3.95±0.1	-1.09c	-0.22c	+0.12c	-0.40c	-3.97±0.16
m(U/M)	2.23±0.11	2.23d	2.23d	2.23d	2.23d	2.08±0.08
Apo oxidizeda, e
$\mathrm{\Delta }{\text{G}}_{(\text{U}/\text{M})}^{°}$ b	-4.48±0.07	-2.83±0.07	-2.32±0.15	-1.87±0.07	-2.52±0.08	-4.54±0.10
m(U/M)	1.65±0.02	1.88±0.04	1.91±0.08	1.89±0.04	2.09±0.05	1.73±0.03
Apo oxidizeda, e
kf	0.056±0.002	0.15±0.02	0.033±0.006	0.029±0.004	0.030±0.003	0.039±0.004
$m_f^{‡}$	1.20±0.05	1.5±0.1	1.36±0.20	1.37±0.18	1.33±0.13	1.13±0.04
ku	(2.02±0.40)×10-5	(1.0±0.1)×10-3	(3.5±1.5)×10-4	(1.2±0.2)×10-3	(5.0±1.0)×10-4	(1.2±0.1)×10-5
$m_u^{‡}$	-0.58±0.01	-0.64±0.03	-0.78±0.09	-0.74±0.03	-0.74±0.05	-0.65±0.01
$\mathrm{\Delta }{\text{G}}_{(U/M)}^{°}$	-4.61±0.12	-2.92±0.2	-2.65±0.28	-1.85±0.13	-2.37±0.13	-4.70±0.08
m(U/M)f	1.79±0.05	2.14±0.1	2.14±0.22	2.11±0.18	2.07±0.14	1.78±0.04
Zn oxidizedg
kf	0.74±0.01	0.87±0.15	0.27±0.05	0.19±0.05	0.34±0.05	0.64±0.01
$m_f^{‡}$	2.93±0.07	3.0±0.1	2.89±0.09	2.48±0.17	3.39±0.10	2.95±0.10
ku	(0.23±0.08)×10-7	(1.77±0.39)×10-7	(2.8±0.8)×10-7	(4.1±1.2)×10-7	(13.3±3.9)×10-7	(2.8±1.2)×10-7
$m_u^{‡}$	-1.76±0.05	-1.85±0.03	-2.01±0.06	-1.88±0.06	-1.68±0.06	-1.47±0.13
$\mathrm{\Delta }{\text{G}}_{(U/M)}^{°}$ h	-18.0±0.2	-17.0±0.2	-15.3±0.5	-15.6±0.2	-15.2±0.2	-16.5±0.3
m(U/M)f	4.69±0.09	4.85±0.1	4.90±0.11	4.36±0.18	5.07±0.12	4.42±0.13
$\mathrm{\Delta }\mathrm{\Delta }{G}_{Zn}^{°}$	-13.4±0.2	-14.1±0.2	-12.7±0.5	-13.8±0.2	-12.8±0.2	-11.8±0.3
Kdh	100±39 pM	31±15 pM	360±350 pM	55±23 pM	270±110 pM	1600±840 pM

^*Units for the U ⇆ M reaction: k_f and k_u, s^-1; $m_f^{‡}$, $m_u^{‡}$ and m_(U/M), kcal mol^-1 M^-1; $\mathrm{\Delta }{\text{G}}_{(\text{U}/\text{M}.)}^{°}$.

$\mathrm{\Delta }\mathrm{\Delta }{G}_{Zn.}^{°}$. kcal mol^-1.

^aDenaturation was performed with urea.

^bEquilibrium data were fit to a two-state model, U⇆M, by globally analyzing the CD signal at 20 different wavelengths between 220 and 240 nm.

^cDue to the constraints required to fit these titrations, the error in the free energy of folding is approximated to be ±0.3 kcal mol^-1.

^dParameter was fixed to the WT value.

^eWT values adapted from Svensson et al.¹⁶

^fCalculated according to m_tot = |m^‡_u| + |m^‡_f.|

^gDenaturation was performed with Gdn-HCl.

^hThe free energy difference between U+Zn ⇆ N•Zn and the Zn affinity was calculated as described in Kayatekin et al. ³⁰