Scatchard analysis of wild-type and mutant cell lines
| Cell Line | Kd | Bmax |
|---|---|---|
| nM | pmol/mg | |
| CB2 | 0.88 ± 0.09 | 1.55 ± 0.39 |
| K109A | 0.82 ± 0.23 | 1.60 ± 0.21 |
| K109R | 1.68 ± 0.29 | 1.00 ± 0.21 |
| K109AS112G | 3.01 ± 1.10 | 0.47 ± 0.01 |
Saturation experiments using [3H]CP-55,940 were performed on stably transfected HEK-293 cells to evaluate binding affinities and relative levels of receptor expression in the CB2 wild-type and K109A and K109R mutant receptors. Nonspecific binding was determined in the presence of excess CP-55,940 (see Experimental Procedures). [3H]WIN 55,212-2 was used to determineKd and Bmax values for K109AS112G cell line. Data are presented as means ± S.E.M. of three independent experiments, each performed in triplicate.