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. 2019 May 16;116(23):11265–11274. doi: 10.1073/pnas.1821447116

Table 1.

Biophysical properties of Adk mutants

Mutant ΔTm, °C* kcat, μM ADP/min/nM Adk KM, μM AMP KI, μM AMP, Distance from AP5A, Å
V106H −11.7 93.2 (2.1) 133.6 (9.5) NA§ 13.4
V106W −9.9 72.6 (4.6) 195.7 (35.3) NA§ 13.4
L82F −5.7 87.3 (6.9) 84.8 (13.7) 2,302.0 (738.2) 10.5
A93F −5.5 87.0 (8.6) 91.3 (16.5) 889.1 (209.7) 8.7
V106A −5.2 72.0 (6.5) 58.0 (11.1) 1,287.0 (355.5) 13.4
V106L −4.8 106.6 (2.4) 190.5 (12.2) NA§ 13.4
A93L −4.3 80.8 (3.4) 119.8 (16.5) NA§ 8.7
A93Y −3.7 85.7 (7.9) 97.0 (18.0) 3,046.0 (1,349.0) 8.7
A93I −3.6 69.0 (5.6) 349.8 (67.8) NA§ 8.7
L83I −2.5 105.9 (5.4) 259.1 (34.7) NA§ 8.8
WT 0.0 68.6 (3.6) 46.8 (5.2) 930.5 (154.7)
Y182F 0.5 88.1 (7.0) 87.0 (14.0) 2,365.0 (766.8) 10.7
L83F 0.6 73.9 (4.5) 94.1 (9.6) 680.2 (106.1) 8.8
E210L 0.7 85.8 (7.3) 115.1 (15.7) 638.1 (132.1) 13.1
L82V 0.8 28.5 (8.3) 36.2 (17.0) 76.6 (33.4) 10.5
V169E 1.8 63.1 (4.0) 83.6 (9.2) 595.9 (92.9) 10.0
L209I 2.6 36.9 (2.0) 32.7 (3.7) 382.9 (45.0) 9.4
M21A 2.8 68.5 (6.4) 48.2 (9.4) 729.5 (166.3) 12.5
V169E + L209I 2.8 71.8 (3.6) 38.5 (4.0) 460.5 (54.2)
L107I 3.2 23.6 (1.4) 22.4 (3.2) 511.7 (77.4) 9.5
M21A + V169E 3.5 60.4 (2.5) 43.3 (3.6) 456.4 (44.3)
L107I + V169E 3.5 97.6 (5.6) 60.6 (6.3) 453.0 (57.6)
M21A + L209I 3.8 55.7 (2.7) 25.5 (2.6) 227.4 (20.7)
M21A + L107I 4.3 72.2 (5.6) 43.2 (6.0) 211.3 (28.1)
M21A + V169E + L209I 4.7 64.0 (2.5) 31.6 (2.5) 228.9 (16.4)
M21A + L107I + V169E 5.2 68.8 (3.7) 37.7 (4.0) 308.6 (32.9)
Q16Y 5.4 4.9 (1.1) 186.3 (118.3) NA§ 5.1
Q16F 6.5 10.6 (1.4) 324.2 (101.0) NA§ 5.1
Q16F + V169E 7.3 20.6 (3.9) 479.3 (156.5) NA§
Q16F + L107I 8.1 21.9 (2.7) 286.8 (74.2) NA§
Q16F + L107I + V169E 8.5 14.0 (2.8) 241.0 (107.0) NA§
Q16F + M21A 8.7 24.0 (3.4) 491.7 (118.4) NA§
*

Tm of WT Adk is 54.9 °C.

Numbers in parentheses are SEM of minimum three repeats.

The kinetic data were fit to uncompetitive model of inhibition (Methods).

§

The kinetic data were fit to standard Michaelis–Menten equation (Methods).