. 2012 Jan 11;20(1-2):101–112. doi: 10.1016/j.str.2011.11.001

Table 2.

In Vitro Activity of MBP-Ddn and Corresponding nfa33440 Mutants toward PA-824

Ddn Mutant^a	k_cat^b (s⁻¹)	K_M^c (μM)	k_cat/K_M (s⁻¹μM⁻¹)	k_cat/K_M (% WT)	nfa33440 Mutant^a	k_cat^b (s⁻¹)	K_M^c (μM)	k_cat/K_M (s⁻¹μM⁻¹)	k_cat/K_M (%WT)	Binding by NMR^d
WT^e	2.2 (±0.3)	15.5 (±0.06)	0.14 (±0.02)	100	WT^e	0.33 (±0.08)	21.9 (±8.5)	0.016 (±0.002)	100
F16L	2.0	30	0.07	47
F16V	2.2	37	0.06	43
F16A	2.3	37	0.06	44
F17L	2.3	18	0.13	93
F17V	2.5	35	0.07	51
F17A	2.1	22	0.10	68
F16/F17A	2.3	45	0.05	37
F16/F17/W20A^e	1.2 (±0.01)	104.1 (±4.0)	0.01 (±0.001)	8
W20F	2.5	20	0.13	93
W20L	2.6	36	0.07	51
W20A	2.7	23	0.12	86	W16A	0.2	17	0.012	75	No change
S22A	1.5	15	0.1	71
S22V	1.3	34	0.04	26
F41L	1.9	14	0.13	93
F41A	1.3	26	0.05	36	F35A	<0.1				PA-824 weaker
Q42L	1.2	16	0.08	54
Y65F^e	1.1 (±0.04)	10.4 (±1.7)	0.10 (±0.01)	71
Y65L	<0.1
Y65A	<0.1				M59A	<0.1				F₄₂₀, PA-824 weaker
A76G	<0.1				V70A	<0.2
S78A	<0.1				S72A	<0.1
					S72V	<0.1				PA-824 stronger
K79L	<0.1				Q73A	<0.2				No change
Y130F^e	0.8 (±0.02)	24.8 (±1.8)	0.03 (±0.004)	21	Y124F	0.28	43	0.007	41
Y130L	<0.1				Y124L	<0.1
Y130A	<0.1
S132V	2.5	52	0.05	34	T126V	0.22	18	0.012	75	No change
S132A	2.1	19	0.11	79
Y133F	<0.1				Y127F	0.23	85	0.003	17
Y133L	<0.1				Y127L	<0.1
Y133A	<0.1
D135N	2.7	19	0.14	100	E129A	0.43	74	0.006	36	No change
Y136F^e	2.2 (±0.6)	13.1 (±2.7)	0.16 (±0.02)	114	Y130F	0.31	26	0.012	75
Y136V	<0.1
Y136L	<0.1				Y130L	<0.1
R142A	<0.1				R136A	<0.1				PA-824 weaker
R142L	<0.1				R136Q	<0.1				PA-824 weaker
I144V	1.7	17	0.10	69
I144A	<0.1				I138A	<0.1				PA-824 weaker
I144G	<0.1

See Figure S7 for the locations of all tested mutants.

For mutants with k_cat values <0.2, the K_M (and hence k_cat/K_M) could not be determined because of the very low reaction rates under the assay conditions.

K_M for PA-824 at saturating [F₄₂₀].

See Figures S5 and S6 and Tables S3 and S4 for additional information.

Means with standard errors from triplicate experiments are reported for some proteins.