Table 1. cAMP concentration and PKA activity in different compartments of healthy and hypertrophic myocytes.
|
Control |
Hypertrophic |
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|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Stimuli |
1 nM ISO |
0.5 nM ISO |
100 μM IBMX |
0.5 nM ISO |
||||||||||||
| cAMP | PKA | cAMP | PKA | cAMP | PKA | cAMP | PKA | |||||||||
| Compartments | ||||||||||||||||
| AKAP18δ | 5.8 | 45 | 3 | 13 | 5.1 | 37 | 2.6 | 9 | ||||||||
| TPNI | 3.7 | 21 | 2.1 | 5 | 5 | 36 | 1.4 | 0 | ||||||||
| AKAP79 | 5.4 | 40 | 3.3 | 16 | 4 | 35 | 2.8 | 11 | ||||||||
| Bulk cytosol | 4.1 | 26 | 2.6 | 9 | 3 | 14 | 1.6 | 2 | ||||||||
Concentration of cAMP (μM) and increase in PKA activity over basal (expressed as percent of maximal activation) in untreated (control) and NE-treated (hypertrophic) NRVM at AKAP18δ, TPNI, AKAP79 and in the bulk cytosol. cAMP concentrations are calculated from FRET changes as measured with AKAP18δ-CUTie, TPNI-CUTie, AKAP79-CUTie or CUTie using the calibration curves shown in Figs 1e and 2f. FRET changes to calculate cAMP concentrations are from Supplementary Fig. 4b,e and Fig. 6b,c. For completeness, data for cAMP changes in the bulk cytosol in response to 1 nM ISO and 100 μM IBMX are also shown. Increase in PKA activity is calculated from cAMP concentrations derived as above using the PKA activity calibration curve shown in Supplementary Fig. 9. Basal PKA activity is set at 3% based on the in-cell PKA activity calibration curve.