Allosteric ligand efficacy (logτB(C)) and functional cooperativity (logαβ) for WT and mutant 3xHA-A1ARs determined from functional interaction studies in cAMP accumulation assays
Values represent the mean ± S.E.M. from indicated number of separate experiments (n) performed in duplicate.
| logτB(C)a | logαβb (αβ) | |||
|---|---|---|---|---|
| PD81723 | VCP171 | PD81723 | VCP171 | |
| WT | −0.13 ± 0.09 (n = 8) | −0.26 ± 0.09 (n = 9) | 0.63 ± 0.05 (4.3) | 0.61 ± 0.06 (4.1) |
| F144A | −0.37 ± 0.08 (n = 3) | −0.59 ± 0.21 (n = 3) | 0.65 ± 0.06 (4.4) | 0.47 ± 0.08 (2.9) |
| G145A | −0.70 ± 0.35 (n = 3) | −0.74 ± 0.09 (n = 3) | 0.64 ± 0.08 (4.4) | 0.55 ± 0.10 (3.5) |
| W146A | −0.62 ± 0.27 (n = 3) | −0.35 ± 0.12 (n = 3) | 0.57 ± 0.04 (3.7) | 0.50 ± 0.05 (3.2) |
| N147A | −0.32 ± 0.16 (n = 3) | −0.28 ± 0.17 (n = 3) | 0.38 ± 0.13 (2.4) | 0.62 ± 0.09 (4.2) |
| N148A | −0.80 ± 0.21 (n = 3)* | −1.24 ± 0.36 (n = 3)* | 0.54 ± 0.06 (3.5) | 0.36 ± 0.07 (2.3) |
| L149A | −0.40 ± 0.2 (n = 3) | −0.31 ± 0.07 (n = 3) | 0.33 ± 0.11 (2.1) | 0.33 ± 0.05 (2.1) |
| SV152AA | −0.10 ± 0.20 (n = 3) | −0.14 ± 0.13 (n = 3) | 0.84 ± 0.10 (6.9) | 0.53 ± 0.05 (3.4) |
| E153A | −0.78 ± 0.08 (n = 3)* | −1.37 ± 0.12 (n = 3)* | 0.67 ± 0.18 (4.7) | 0.59 ± 0.09 (3.9) |
| R154A | −0.63 ± 0.21 (n = 3) | −1.04 ± 0.18 (n = 3)* | 0.63 ± 0.03 (4.3) | 0.26 ± 0.02 (1.8)* |
| W156A | −0.31 ± 0.19 (n = 3) | −0.66 ± 0.18 (n = 3) | 0.66 ± 0.05 (4.5) | 0.29 ± 0.06 (2.0)* |
| N159A | −0.56 ± 0.14 (n = 3) | −0.72 ± 0.04 (n = 3) | 0.44 ± 0.08 (2.8) | 0.25 ± 0.04 (1.8)* |
| G160A | −0.43 ± 0.09 (n = 3) | −0.46 ± 0.16 (n = 3) | 0.58 ± 0.07 (3.8) | 0.51 ± 0.04 (3.2) |
| S161A | −0.85 ± 0.29 (n = 3)* | −1.21 ± 0.59 (n = 3)* | 0.32 ± 0.06 (2.1) | 0.33 ± 0.03 (2.1) |
| MGE162AAA | −0.43 ± 0.30 (n = 3) | −0.15 ± 0.13 (n = 3) | 0.55 ± 0.19 (3.5) | 0.49 ± 0.17 (3.1) |
| P165A | −0.07 ± 0.08 (n = 4) | −0.28 ± 0.01 (n = 3) | 0.59 ± 0.05 (3.9) | 0.46 ± 0.07 (2.9) |
| V166A | −0.27 ± 0.09 (n = 4) | −0.40 ± 0.13 (n =3) | 0.59 ± 0.04 (3.9) | 0.42 ± 0.08 (2.6) |
| I167A | −0.78 ± 0.17 (n = 4)* | −1.07 ± 0.30 (n = 3)* | 0.49 ± 0.08 (3.1) | 0.38 ± 0.01 (2.4) |
| K168A | −0.11 ± 0.05 (n = 4) | −0.29 ± 0.12 (n = 3) | 0.49 ± 0.07 (3.1) | 0.48 ± 0.14 (3.0) |
| E170A | −0.43 ± 0.13 (n = 4) | −0.60 ± 0.14 (n = 3) | 0.51 ± 0.07 (3.3) | 0.42 ± 0.12 (2.6) |
| E172A | −0.39 ± 0.29 (n = 3) | −0.63 ± 0.04 (n = 3) | 0.95 ± 0.18 (8.9)* | 0.62 ± 0.04 (4.2) |
| K173A | −0.60 ± 0.21 (n = 3) | −0.53 ± 0.27 (n = 3) | 0.33 ± 0.08 (2.1) | 0.38 ± 0.13 (2.4) |
| V174A | −0.43 ± 0.04 (n = 4) | −0.66 ± 0.29 (n = 3) | 0.53 ± 0.04 (3.4) | 0.27 ± 0.07 (1.8)* |
| I175A | −0.74 ± 0.06 (n = 3) | −1.42 ± 0.37 (n = 3)* | 0.33 ± 0.10 (2.1) | 0.24 ± 0.02 (1.7)* |
| S176A | −0.36 ± 0.11 (n = 4) | −0.31 ± 0.14 (n = 3) | 0.57 ± 0.03 (3.7) | 0.31 ± 0.07 (2.0) |
| M177A | −0.52 ± 0.08 (n = 4) | −0.23 ± 0.08 (n = 3) | 0.58 ± 0.03 (3.8) | 0.44 ± 0.07 (2.8) |
↵a Logarithm of the efficacy parameter determined from the operational model of allosterism corrected for changes in receptor expression.
↵b Logarithm of the functional cooperativity between the allosteric ligand and NECA. Antilogarithm is shown in parentheses.
↵* P < 0.05, significantly different from WT, one-way analysis of variance, Dunnett’s post hoc test.