. Author manuscript; available in PMC: 2018 Sep 28.

Published in final edited form as: J Am Chem Soc. 2017 Mar 28;139(14):5051–5060. doi: 10.1021/jacs.6b11323

Table 1.

Thermodynamic and kinetic parameters for engrailed-HD and consensus-HD.

	Folding thermodynamics		Folding kinetics				Binding thermodynamics

	ΔG°(H₂O)(kcal·mol⁻¹)	m-value (kcal·mol⁻¹·M⁻¹)	$k_{f, H_{2} O}$ (s⁻¹)	m_f (Μ⁻¹)	$k_{u, H_{2} O}$ (s⁻¹)	m_u (Μ⁻¹)	n	K_d (nM)	ΔΗ° (kcal·mol⁻¹)	TΔS° (kcal·mol⁻¹)

Engrailed-HD	3.62 ± 0.03	1.39 ±0.01	3.9 × 10⁴	−0.81	67.1	0.17	0.98 ± 0.01	737 ± 38	−9.2 ±0.1	−0.8 ± 0.1
Consensus-HD	8.05 ±0.10	1.45 ± 0.02	1.1 × 10^e	−0.83	1.4	NA^a	1.00 ±0.01	8.1 ±1.6	−16 ±0.2	−5.0 ± 0.3
Consensus-HD (10°C)	8.51 ± 0.27	1.35 ± 0.02	5.50 × 10	−0.90	3.24	0.23	ND	ND	ND	ND

Folding thermodynamic: parameters were obtained by fitting gaunidine-induced denaturation curves. Uncertainies are standard errors on the mean of three titrations. Conditions: 25 mM NaPO₄ (pH 7.0), 150 mM NaCI, 20°C

Folding kinetics: Parameters were obtained by fitting equation (2) to the guanidine dependence of the rate constants for the refolding and unfolding 1 phases (Figure 5B, C).

For consensus-HD, the m_u parameter was fixed to the fitted value for the engrailed-HD (0.17 M⁻¹). Conditions: 25 mM NaPO₄, 150 mM NaCI, temperatures as indicated.

Binding thermodynamics: Data were anaylzed using Origin 7.0 software fitting to a single site model. Uncertainies are standard errors on the mean of three independent experiments Conditions: 25 mM HEPES (pH 7.5), 250 mM KCI, 25°C.