Treatment | Construct | |||
---|---|---|---|---|
5-HT1A(Cys351Ile)Gi1α | 5-HT1A(Cys351Ile)Go1α | |||
Vmax | Km | Vmax | Km | |
pmol GTP/min/mg protein | nM | pmol GTP/min/mg protein | nM | |
Basal | 33.1 ± 2.7 | 169 ± 12 | 29.5 ± 2.5 | 158 ± 6 |
5-HT | 58.0 ± 6.6 | 217 ± 9 | 42.9 ± 6.0 | 136 ± 11 |
WAY100635 | 39.7 ± 4.2 | 207 ± 14 | 35.9 ± 4.6 | 178 ± 16 |
RGS1 | 56.4 ± 2.1 | 300 ± 6 | 151 ± 2 | 504 ± 23 |
RGS1 +5-HT | 313 ± 15 | 1174 ± 38 | 644 ± 9 | 1353 ± 160 |
RGS1 +WAY100635 | 66.8 ± 3.1 | 304 ± 9 | 192 ± 17 | 576 ± 54 |
Enzyme kinetic analysis was performed on high-affinity GTPase activity and its regulation in membranes of HEK293 cells expressing either the 5-HT1A(Cys351Ile)Gi1α or 5-HT1A(Cys351Ile)Go1α fusion proteins. Data are taken from representative experiments. RGS1 and the ligands were present at 1 μM.