Arterioscler Thromb Vasc Biol. 2011 Oct;31(10):2278-86. doi: 10.1161/ATVBAHA.110.221010. Epub 2011 Jul 28.
Cilostazol suppresses angiotensin II-induced vasoconstriction via protein kinase A-mediated phosphorylation of the transient receptor potential canonical 6 channel.
Nishioka, K., Nishida, M., Ariyoshi, M., Jian, Z., Saiki, S., Hirano, M., Nakaya, M., Sato, Y., Kita, S., Iwamoto, T., Hirano, K., Inoue, R., Kurose, H.,
["Department of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan."]
["Department of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan."]
OBJECTIVE: The goal of this study was to determine whether inhibition of transient receptor potential canonical (TRPC) channels underlies attenuation of angiotensin II (Ang II)-induced vasoconstriction by phosphodiesterase (PDE) 3 inhibition. METHODS AND RESULTS: Pretreatment of rat thoracic aorta with cilostazol, a selective PDE3 inhibitor, suppressed vasoconstriction induced by Ang II but not that induced by KCl. The Ang II-induced contraction was largely dependent on Ca(2+) influx via receptor-operated cation channels. Cilostazol specifically suppressed diacylglycerol-activated TRPC channels (TRPC3/TRPC6/TRPC7) through protein kinase A (PKA)-dependent phosphorylation of TRPC channels in HEK293 cells. In contrast, we found that phosphorylation of TRPC6 at Thr69 was essential for the suppression of Ang II-induced Ca(2+) influx by PDE3 inhibition in rat aortic smooth muscle cells (RAoSMCs). Cilostazol specifically induced phosphorylation of endogenous TRPC6 at Thr69. The endogenous TRPC6, but not TRPC3, formed a ternary complex with PDE3 and PKA in RAoSMCs, suggesting the specificity of TRPC6 phosphorylation by PDE3 inhibition. Furthermore, inhibition of PDE3 suppressed the Ang II-induced contraction of reconstituted ring with RAoSMCs, which were abolished by the expression of a phosphorylation-deficient mutant of TRPC6. CONCLUSIONS: PKA-mediated phosphorylation of TRPC6 at Thr69 is essential for the vasorelaxant effects of PDE3 inhibition against the vasoconstrictive actions of Ang II.
PMID: 21799177
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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Screening
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| Experimental screening | Non-experimental screening | Reference | ||||||||
| TRP channel construct | Interactor source | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Organ/tissue | Sample type | |||
| TRPC3 |
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PDE3A | Inference | Prediction | 21799177 | |||||
| TRPC3 |
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PKA | Inference | Prediction | 21799177 | |||||
| TRPC6 |
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PDE3A | Inference | Prediction | 21799177 | |||||
| TRPC6 |
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PKA | Inference | Prediction | 21799177 | |||||
(:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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top
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| Validation: In vivo validation
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| Assay with endogenous proteins | Assay with overexpressed proteins | Reference | ||||||||
| Cell or tissue | Cell or tissue | TRP channel construct | Interactor construct | |||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPC3 |
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PDE3A | Co-immunoprecipitation | HEK293 | Not specified | Full-length | Not used | 21799177 | ||
| TRPC3 |
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PKA | Co-immunoprecipitation | HEK293 | Not specified | Full-length | Not used | 21799177 | ||
| TRPC6 |
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PDE3A | Co-immunoprecipitation | HEK293 | Mouse | Full-length | Not used | 21799177 | ||
| TRPC6 |
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PDE3A | Co-immunoprecipitation | Rat aortic smooth muscle cell | 21799177 | |||||
| TRPC6 |
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PKA | Co-immunoprecipitation | HEK293 | Mouse | Full-length | Not used | 21799177 | ||
(:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)