A Membrane Permeable Prodrug of S223 for Selective Epac2 Activation in Living Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. NMR
2.2. Hydrolysis of S223-AM
2.3. U2OS Cell Model System
2.4. Insulin-Secreting MIN6 β-cells
2.5. Pancreatic Islets
2.6. Total Internal Reflection Fluorescence Imaging of Epac Translocation and Plasma Membrane Activities of Rap and PKA
3. Results
3.1. Synthesis of S223-AM
3.2. Stability and Hydrolysis Mechanism of S223-AM
3.3. Characterisation of S223-AM in Living Cells
3.4. S223-AM Selectively Activates Epac2 in β-cells
3.5. S223-AM Promotes Rap Activation at the β-Cell Plasma Membrane
3.6. S223-AM Does Not Activate PKA in β-Cells
3.7. cAMP-Dependent Rap Activity in β-cells is Mediated by Epac2
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Atom | 31P | 13C | 1H | ||
---|---|---|---|---|---|
/ppm | /ppm | /ppm | Multi-plicity | J/Hz | |
P | 19.92 | ||||
2 | 155.2 | 8.202 | s | ||
4 | 153.5 | ||||
5 | 122.5? | ||||
6 | 157.6 | ||||
8 | 150.1 | ||||
6N’ | 7.330 | ||||
6N’’ | 6.788 | ||||
1’ | 92.75 | 6.001 | s | ||
2’ | 83.31 | 4.73 | d | 5.3 | |
3’ | 81.06 | 5.752 | m | 5.1/5.3 | |
4’ | 73.30 | 4.335 | td | 4.9/10.3 | |
5’ | 73.88 | 4.828 | dq | 4.9/23.3 | |
5” | 4.412 | t | 10.1 | ||
L1 | 39.50 | 4.679 | d | 3.2 | |
L2 | 140.1 | ||||
L3 | 132.3 | 7.561 | d | 8.0 | |
L4 | 131.52 | 7.422 | t | 7.3 | |
L5 | 130.66 | 7.377 | t(d) | 7.3 (2.0) | |
Z1 | 61.37 | 3.521 | s | ||
A1 | 23.9 | 2.177 | s | ||
A2 | 172.8 | ||||
A3′ | 65.0 | 5.653 | dd | 11.2/21.2 | |
A3′’ | 5.571 | dd | 11.2/18.8 |
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Xu, Y.; Schwede, F.; Wienk, H.; Tengholm, A.; Rehmann, H. A Membrane Permeable Prodrug of S223 for Selective Epac2 Activation in Living Cells. Cells 2019, 8, 1589. https://doi.org/10.3390/cells8121589
Xu Y, Schwede F, Wienk H, Tengholm A, Rehmann H. A Membrane Permeable Prodrug of S223 for Selective Epac2 Activation in Living Cells. Cells. 2019; 8(12):1589. https://doi.org/10.3390/cells8121589
Chicago/Turabian StyleXu, Yunjian, Frank Schwede, Hans Wienk, Anders Tengholm, and Holger Rehmann. 2019. "A Membrane Permeable Prodrug of S223 for Selective Epac2 Activation in Living Cells" Cells 8, no. 12: 1589. https://doi.org/10.3390/cells8121589