Table 2.

Mutations validation on the ligand binding pocket using thermostability and affinity MS assay

Mutants	(–)-Crebanine/CPM (n = 2) (°C)	(–)-Crebanine/MS (n = 2)a (%)
WT	59.93	27.57
G2185.42A	53.92	9.82
G2185.42S	53.24	6.04
A2225.46V	59.55	25.84
A2225.46F	55.70	15.87
T1393.37A	54.63	14.64
L209ECL2F	55.46	13.22

^aRelative ligand binding (%) was calculated by the MS response of the ligand released from a specific mutant divided by the MS response of the total ligand present in the incubation sample