Table 2.

Enzyme kinetic constants and protein melting temperatures for NS3-4A protease variants

Kinetic constants (left panel) and melting temperatures (right panel) were obtained by a FRET-based protease and thermal shift assay, respectively. Data shown represent the mean ± S.D. from at least three independent experiments.

Substrate	Variant	Enzyme kinetics			Variant	Melting temperature Tm (°C)
		Km (μm)	kcat (/s)	kcat/Km (1/(s*µm))
NS4A/4B	WT (Q41)	3.11 ± 0.29	0.19 ± 0.011	0.062 ± 0.008	WT (Q41)	54.9 ± 0.3
	Q41R	2.11 ± 0.14	0.18 ± 0.006	0.085 ± 0.008	Q41R	55.1 ± 0.3
	D168A	2.45 ± 0.23	0.09 ± 0.004	0.035 ± 0.004	D168A	48.7 ± 0.1
	Q41R-D168A	1.71 ± 0.23	0.15 ± 0.009	0.087 ± 0.015	Q41R-D168A	48.3 ± 0.2
	D168E	1.79 ± 0.11	0.12 ± 0.003	0.067 ± 0.005	D168E	55.9 ± 0.2
	Q41R-D168E	1.83 ± 0.10	0.14 ± 0.001	0.074 ± 0.005	Q41R-D168E	55.8 ± 0.3
	D168T	2.03 ± 0.15	0.13 ± 0.005	0.064 ± 0.006	D168T	47.1 ± 0.1
	Q41R-D168T	1.69 ± 0.14	0.12 ± 0.004	0.071 ± 0.007	Q41R-D168T	48.5 ± 0.2
MAVS	WT (Q41)	7.82 ± 0.70	0.058 ± 0.002	0.008 ± 0.001
	Q41R	37.32 ± 4.42	0.275 ± 0.021	0.007 ± 0.001
	D168A	14.41 ± 2.91	0.029 ± 0.003	0.002 ± 0.000
	Q41R-D168A	1.77 ± 0.14	0.507 ± 0.011	0.287 ± 0.024