B2 Receptor c-myc Tagged | Kd[3H]BK | |||
---|---|---|---|---|
Intact Cells | Membrane | |||
No Pretreatment | Na+ Pretreatment | Na+ + GTPγS Pretreatment | ||
nM | ||||
WT | 0.54 ± 0.17 | 0.47 ± 0.17 | 2.2 ± 0.45 | 3.7 ± 0.7 |
B max = 0.49 | B max = 0.39 | B max = 0.38 | ||
pmol/mg | pmol/mg | pmol/mg | ||
N113A | 0.67 ± 0.22 | 0.21 ± 0.11 | 0.28 ± 0.15 | 0.27 ± 0.07 |
B max = 1.1 | B max = 1.0 | B max = 1.1 | ||
pmol/mg | pmol/mg | pmol/mg | ||
D76N | 0.83 ± 0.30 | 0.51 ± 0.15 | 2.5 ± 0.4 | N.D.1-a |
B max = 1,0 | B max = 0,81 | |||
pmol/mg | pmol/mg | |||
W256F | 0.52 ± 0.18 | N.D. | N.D. | N.D. |
W256Q | 0.41 ± 0.15 | N.D. | N.D. | N.D. |
Y295F | 0.31 ± 0.13 | N.D. | N.D. | N.D. |
The parameters of [3H]BK binding to intact COS-7 cells expressing comparable levels of the WT or mutant receptors, or to membrane preparations from these cells, were carried out as described in Materials and Methods. The reported K dvalues are the mean ± S.D. of three independent experiments, carried out with triplicate assays. The B max values are those of a typical experiment involving comparison of WT and mutant receptors expressed at similar levels.
↵1-a N.D., not determined.