Table 2.

Changes in the selectivity of APPI-3M variants with mutations at selectivity-switch residues toward human serine proteases

Mutation	Target A	Target B	Predicted switcha	Ki [pM], target Ab	Ki [pM], target Bb	Switch ratioc
T11H	Mesotrypsin	Anionic trypsin	A –>B	302 ± 24	4.45 ± 0.23	1.6
T11I			B –>A	61.2 ± 2.6a	1.44 ± 0.14
Ki Fold changed	—	—	—	4.93	3.09	—
G17E	KLK6	Cationic trypsin	A –>B	464 ± 39	85.8 ± 5 .9	5.7
G17R			B –>A	77.4 ± 2.6	8.12 ± 0.08
Ki Fold changee	—	—	—	59.95	10.57	—

^aAs predicted by the NGS analysis

^bResults (means ± SD) were obtained from three independent experiments

^cCalculated as the K_i fold change for target A divided by the K_i fold change for target B

$Switchratio=\frac{K_{i}foldfortargetA}{K_{i}foldfortargetB}$

^dK_i (T11H)/K_i (T11I)

^eK_i (G17E)/K_i (G17R)