Hypothetical protein CuvA (Rv1422) from Mycobacterium tuberculosis H37Rv interacts with uridine diphosphate N-acetylglucosamine as a key precursor of cell wall

Applied Biological Chemistry

Table 3 Kinetic parameters for the binding of UDP-GlcNAc to MtCuvA mutants

UDP-GlcNAc	K_D (mM)	ΔT_m max	Binding efficiency (ΔT_m max/K_D)	R²
Wild type	0.38 ± 0.06	9.47 ± 0.34	24.92 ± 4.04	0.986
H12A	0.69 ± 0.11	7.30 ± 0.32	10.58 ± 1.75	0.986
T33A	1.77 ± 0.24	5.63 ± 0.28	3.18 ± 0.46	0.992
D36A	0.56 ± 0.09	7.24 ± 0.32	12.93 ± 2.15	0.984
R80A	0.50 ± 0.07	9.44 ± 0.31	19.07 ± 2.61	0.990
Q154A	2.48 ± 0.61	4.02 ± 0.41	1.62 ± 0.43	0.976
S196A	1.45 ± 0.14	7.25 ± 0.24	5.01 ± 0.51	0.996
T199A	5.25 ± 1.18	4.66 ± 0.55	0.89 ± 0.23	0.988
S200A	0.20 ± 0.03	10.47 ± 0.30	52.35 ± 7.99	0.988
N226A	0.98 ± 0.09	8.90 ± 0.25	9.08 ± 0.87	0.996
E230A	0.26 ± 0.04	9.38 ± 0.28	36.08 ± 5.65	0.998
T295A	0.37 ± 0.05	9.20 ± 0.29	24.86 ± 3.45	0.990
H298A	ND^*	ND	ND	−

R² = Coefficient of determination
^*ND means “no determined” and indicates that the binding of UDP-GlcNAc to MtCuvA and the Δ T_m values are too weak to calculate the K_D