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. 2016 Feb 29;113(11):E1470–E1478. doi: 10.1073/pnas.1601441113

Table S1.

In vitro properties determined for DHFR mutants

DHFR variant kcat, s−1 Km, μM kcat/Km, s−1⋅μM−1 Ki, nM Bis-ANS Tm, °C ΔTm, °C
E. coli
 WT 14 0.87 16.4 0.94 1.0 57.7 0
 P21L 9.3 0.34 27.3 1.3 1.2 49.3 −8.4
 A26T 5.4 0.78 6.97 8.0 2.3 52.9 −4.8
 L28R 1.7 0.22 7.70 29 0.4 63.7 6
 P21L/A26T 7.9 0.94 8.38 12 2.1 43.7 −14
 P21L/L28R 0.31 0.14 2.29 162 0.9 57.2 −0.5
 A26T/L28R 1.6 0.19 8.39 98 1.7 58.1 0.4
 P21L/A26T/L28R 1.0 0.30 3.14 242 2.1 58.5 0.8
L. grayi
 WT 67 1.80 37.1 1.6 7.9 53.4 −4.3
 P21L 5.6 0.38 14.8 3.9 8.0 50.7 −7
 A26T 55 1.76 30.9 2.2 7.4 49.8 −7.9
 L28R 7.2 2.56 2.80 19 5.3 50.1 −7.6
 P21L/A26T 15 5.50 2.70 25 5.8 48.5 −9.2
 P21L/L28R 0.2 0.4 0.48 152 12.1 51.2 −6.5
 A26T/L28R 0.2 0.2 0.91 140.0 8.8 52.5 −5.2
 P21L/A26T/L28R 0.15 0.27 0.58 335 5.7 47.9 −9.8
C. muridarum
 WT 11 5.48 2.10 1.6 6.1 56.4 −1.3
 P23L 5.4 2.67 2.02 4.9 5.8 58.2 0.5
 E28T 5.1 1.65 3.11 2.2 8.9 55 −2.7
 L30R 1.0 1.27 0.77 100 3.8 58.4 0.7
 P23L/E28T 1.2 0.66 1.75 4.7 7.9 58.1 0.4
 P23L/L30R 0.24 0.65 0.37 76 3.7 57.6 −0.1
 E28T/L30R 1.3 0.76 1.73 40 5.1 55.1 −2.6
 P23L/E28T/L30R 0.61 1.00 0.61 92 7.5 55.4 −2.3