VOLUME 291 (2016) PAGES 21925–21944
PAGES 21931 AND 21933:
There were some errors in Tables 3 and 4 whereby some data values were increased by the addition of 10 units to each data point. These errors have now been corrected and do not affect the results or conclusions of this work.
TABLE 3.
Potency (pEC50), affinity (pKa) and coupling efficacy (log τ) values for cAMP production at the CLR co-expressed with each RAMP, stimulated with various agonists measured in HEK-293 cells in the presence and absence of pertussis toxin
Data are the mean ± S.E. of n individual data sets. Statistically different between PTX-treated and untreated was determined using Student's t test (*, p < 0.05; **, p < 0.01; ***, p < 0.001; and ****, p < 0.0001).
| Untreated |
Treated |
|||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| pEC50a | Emaxb | pKac | log τd | n | pEC50a | Emaxb | pKac | log τd | n | |
| RAMP1 | ||||||||||
| CGRP | 9.66 ± 0.2 | 47.07 ± 2.2 | 9.43 ± 0.2 | −0.11 ± 0.04 | 9 | 9.65 ± 0.2 | 44.95 ± 2.2 | 9.33 ± 0.3 | −0.11 ± 0.07 | 6 |
| AM | 7.93 ± 0.2 | 48.06 ± 2.5 | 7.67 ± 0.2 | −0.09 ± 0.05 | 9 | 8.14 ± 0.07 | 72.17 ± 1.7*** | 7.66 ± 0.2 | 0.36 ± 0.1** | 6 |
| AM2 | 7.93 ± 0.2 | 46.10 ± 4.1 | 7.70 ± 0.2 | −0.11 ± 0.07 | 9 | 9.15 ± 0.1* | 72.15 ± 2.4*** | 8.56 ± 0.3 | 0.4 ± 0.1** | 6 |
| RAMP2 | ||||||||||
| CGRP | 9.00 ± 0.2 | 36.97 ± 2.4 | 8.82 ± 0.2 | −0.27 ± 0.05 | 9 | 8.25 ± 0.4 | 56.27 ± 1.4*** | 7.92 ± 0.2* | 0 0.1 ± 0.06** | 6 |
| AM | 10.35 ± 0.1 | 56.33 ± 1.6 | 10.00 ± 0.1 | 0.07 ± 0.02 | 9 | 10.16 ± 0.07 | 56.07 ± 1.1 | 9.83 ± 0.2 | 0.07 ± 0.02 | 6 |
| AM2 | 7.46 ± 0.2 | 36.61 ± 3.5 | 7.24 ± 0.2 | −0.29 ± 0.07 | 9 | 9.13 ± 0.1** | 56.05 ± 2.2*** | 8.84 ± 0.2** | 0.1 ± 0.06* | 6 |
| RAMP3 | ||||||||||
| CGRP | 7.75 ± 0.3 | 22.38 ± 2.6 | 7.64 ± 0.3 | −0.54 ± 0.07 | 8 | 8.90 ± 0.1* | 32.61 ± 1.5* | 8.74 ± 0.2* | −0.29 ± 0.06 | 7 |
| AM | 8.98 ± 0.2 | 32.00 ± 1.5 | 8.83 ± 0.1 | −0.33 ± 0.03 | 8 | 9.10 ± 0.2 | 35.95 ± 2.2 | 8.94 ± 0.2 | −0.34 ± 0.05 | 7 |
| AM2 | 9.10 ± 0.2 | 21.92 ± 1.7 | 9.08 ± 0.2 | −0.51 ± 0.06 | 8 | 8.74 ± 0.2 | 44.35 ± 2.7**** | 8.43 ± 0.1* | −0.07 ± 0.07*** | 7 |
a The negative logarithm of the agonist concentration required to produce a half-maximal response.
b The maximal response to the ligand expressed as a percentage of the maximal cAMP production as determined using 100 μm forskolin stimulation in the presence of pertussis toxin treatment.
c The negative logarithm of the equilibrium disassociation constant for each ligand generated through use of the operational model of agonism (34).
d Log τ is the coupling efficiency parameter of each ligand.
TABLE 4.
Potency (pEC50) and maximal response (Emax), for cAMP production at the CLR co-expressed with each RAMP, stimulated with various agonists measured in HEK-293S cells in the presence or absence of pertussis toxin
Data are the mean ± S.E. of n individual data sets. No statistical difference was found between untreated and PTX-treated HEK-293S cells using Student's t test.
| Untreated |
Treated |
|||||
|---|---|---|---|---|---|---|
| pEC50a | Emaxb | n | pEC50a | Emaxb | n | |
| RAMP1 | ||||||
| CGRP | 9.88 ± 0.1 | 59.98 ± 1.1 | 5 | 9.87 ± 0.1 | 72.92 ± 2.3 | 5 |
| AM | 8.13 ± 0.1 | 60.00 ± 3.1 | 5 | 8.03 ± 0.1 | 61.26 ± 2.6 | 5 |
| AM2 | 8.74 ± 0.1 | 68.94 ± 1.2 | 5 | 8.78 ± 0.1 | 68.30 ± 1.6 | 5 |
| RAMP2 | ||||||
| CGRP | 8.00 ± 0.1 | 32.56 ± 1.0 | 5 | 7.88 ± 0.1 | 39.32 ± 1.7 | 5 |
| AM | 9.39 ± 0.1 | 30.34 ± 0.8 | 5 | 9.38 ± 0.1 | 33.28 ± 1.2 | 5 |
| AM2 | 8.57 ± 0.1 | 40.30 ± 1.5 | 5 | 8.58 ± 0.2 | 30.52 ± 2.0 | 5 |
| RAMP3 | ||||||
| CGRP | 8.42 ± 0.1 | 40.84 ± 0.6 | 5 | 8.38 ± 0.1 | 39.84 ± 1.0 | 5 |
| AM | 9.63 ± 0.1 | 39.09 ± 1.2 | 5 | 9.49 ± 0.2 | 42.26 ± 1.6 | 5 |
| AM2 | 8.01 ± 0.1 | 33.75 ± 1.5 | 5 | 7.79 ± 0.2 | 28.21 ± 2.4 | 5 |
a The negative logarithm of the agonist concentration required to produce a half-maximal response.
b The maximal response to the ligand expressed as a percentage of the maximal cAMP production as determined using 100 μm forskolin stimulation in the presence of PTX treatment.