GnRHR Construct | Receptor Binding | Total IP Production EC50 | |
---|---|---|---|
Kd | Bmax | ||
nM ± S.E.M. | fmol/mg protein ± S.E.M. | nM ± S.E.M. | |
Wild-type | 0.94 ± 0.35 | 541 ± 41 | 2.10 ± 0.20 |
W279S | 2.98 ± 0.89 | 94 ± 19 | Basal |
W279R | ND | ND | Basal |
V299A | ND | ND | Basal |
[W279R/V299A] | 4.95 ± 1.05 | 504 ± 43 | Basal |
[W279R]+[V299A] | ND | ND | Basal |
Receptor binding assays were performed using 125I-GnRH-Ag.K d and B max were calculated from saturation experiments. The half-maximum stimulation (EC50) for total IPs was determined from concentration-dependence incubations. ND, no sufficient binding to allow determination. Basal, absence of GnRH-induced IP production. Values are the mean ± S.E. of at least three individual experiments.