TABLE 2.
Effects of arginine swap mutations on ligand pharmacology at human GPR35
Data are human GPR35a pEC50 values and ΔpEC50 compared with wild-type. All studies employed the BRET-based GPR35–β-arrestin 2 interaction assay. Potency estimates of <4 reflect lack of adequate data fit.
| Ligand | Wild-Type | Arg164Ser |
Arg4.60Met |
Leu4.62Arg |
Arg6.58Gln |
Arg7.32Ser |
Arg6.58Glu, Arg7.32Ser |
||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| pEC50 | ΔpEC50 | pEC50 | ΔpEC50 | pEC50 | ΔpEC50 | pEC50 | ΔpEC50 | pEC50 | ΔpEC50 | pEC50 | ΔpEC50 | ||
| Zaprinast | 5.59 ± 0.01 | 5.45 ± 0.08 | −0.14 | <4 | <−1.59 | 6.13 ± 0.04*** | +0.54 | 5.11 ± 0.05*** | −0.48 | 5.99 ± 0.09*** | +0.4 | 5.08 ± 0.05*** | −0.51 |
| Lodoxamide | 8.44 ± 0.02 | 6.65 ± 0.09*** | −1.79 | 5.88 ± 0.04*** | −2.56 | 8.63 ± 0.09 | +0.19 | 8.19 ± 0.09 | −0.25 | 6.83 ± 0.01*** | −1.61 | <4 | <−4.44 |
| Bufrolin | 7.90 ± 0.04 | 7.36 ± 0.001* | −0.54 | 6.28 ± 0.05*** | −1.62 | 8.18 ± 0.05 | +0.28 | 6.82 ± 0.09*** | −1.08 | 7.61 ± 0.1 | −0.29 | 5.88 ± 0.23*** | −2.02 |
| Cromolyn | 5.20 ± 0.03 | 4.3 ± 0.13*** | −0.9 | <4 | <−1.20 | 5.72 ± 0.04*** | +0.52 | 5.05 ± 0.04 | −0.15 | 5.95 ± 0.15*** | +0.75 | 5.23 ± 0.08 | +0.03 |
| Amlexanox | 5.39 ± 0.06 | 5.28 ± 0.04 | −0.11 | 4.48 ± 0.09*** | −0.91 | 5.83 ± 0.09 | +0.44 | 5.54 ± 0.11 | +0.15 | 5.86 ± 0.15* | +0.47 | 5.74 ± 0.11 | +0.35 |
| Doxantrazole | 5.47 ± 0.07 | 5.27 ± 0.01 | −0.2 | 5.11 ± 0.06 | −0.36 | 5.69 ± 0.06 | +0.22 | 5.09 ± 0.12 | −0.38 | 5.87 ± 0.17 | +0.4 | 5.03 ± 0.11 | −0.44 |
| Pemirolast | <4 | <4 | N/A | <4 | N/A | <4 | N/A | <4 | N/A | <4 | N/A | <4 | N/A |
N/A, not applicable, as pemirolast lacked potency at wild-type human GPR35a.
***
P < 0.001; *P < 0.05: pEC50 significantly different from corresponding wild-type.