TABLE 1

Effects of point mutations in the P-Rex1 PH domain PIP3-binding site on thermal stabilization by compounds 1, 2, and 3

Shown are ΔTm values representing the change in melting temperature of each protein in the presence of compound.

PH variantIP45 μM 120 μM 1100 μM 15 μM 220 μM 25 μM 3
WT10 (6.6, 13)a0.1 (−3.6, 3.8)2.8 (−0.9, 6.5)14 (10, 18)2.7 (−1.1, 6.4)10 (6.5, 14)17 (13, 20)
K280A0.7 (−1.5, 2.9)−0.4 (−2.8, 2)3.5 (1.1, 6)15 (13, 18)1.6 (−0.9, 4)16 (14, 18)18 (16, 21)
S282A5.4 (3.4, 7.4)0.3 (−1.9, 2.5)2.5 (0.3, 4.7)16 (13, 18)1.9 (−0.4, 4.1)12 (9.5, 14)17 (15, 19)
Q287A5.6 (2.7, 8.5)−0.6 (−3.8, 2.6)0.4 (−3.2, 4.1)11 (7.9, 14)2.7 (−0.72, 6.1)9 (5.6, 12)14 (10, 17)
R289A−0.3 (−2.5, 1.8)−0.6 (−3, 1.8)0.9 (−1.4, 3.3)13 (11, 16)3.9 (1.6, 6.3)11 (8.7, 13)16 (13, 19)
Y300F3.8 (1.4, 6.1)−0.7 (−3.3, 1.9)3.3 (0.7, 5.9)16 (13, 18)6 (3, 8.9)16 (14, 19)21 (19, 24)
K302A10 (7, 13)0.1 (−3.2, 3.4)1.6 (−1.7, 4.9)6.6 (3.3, 9.9)2.9 (−0.4, 6.2)11 (7.3, 14)14 (11, 18)
R328A6.1 (3.4, 8.8)−0.6 (−3.3, 2.1)−0.6 (−3.6, 2.4)13 (10, 16)0.8 (−3, 4.6)9.9 (6.6, 13)ND
K368A1.1 (−0.4, 2.6)0.2 (−1.4, 1.9)4.5 (2.9, 6.2)17 (16, 19)3.2 (1.5, 4.8)17 (15, 18)21 (19, 23)
  • ND, not determined; Tm, melting temperature; WT, wild type.

  • a Values shown in parentheses are the 95% confidence intervals.