Cardiovasc Res. 2014 Oct 1;104(1):138-46. doi: 10.1093/cvr/cvu190. Epub 2014 Aug 18.

Nitric oxide and protein kinase G act on TRPC1 to inhibit 11,12-EET-induced vascular relaxation.External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c

Zhang, P., Ma, Y., Wang, Y., Ma, X., Huang, Y., Li, R. A., Wan, S., Yao, X.,
["School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China.", "School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China.", "School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China.", "School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China.", "School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China.", "Stem Cell and Regenerative Medicine Consortium, The University of Hong Kong, Hong Kong, China.", "Department of Surgery, The Chinese University of Hong Kong, Hong Kong, China.", "School of Biomedical Sciences and Li Ka Shing institute of Health Science, The Chinese University of Hong Kong, Hong Kong, China Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China yao2068@cuhk.edu.hk."]
AIMS: Vascular endothelial cells synthesize and release vasodilators such as nitric oxide (NO) and epoxyeicosatrienoic acids (EETs). NO is known to inhibit EET-induced smooth muscle hyperpolarization and relaxation. This study investigates the underlying mechanism of this inhibition. METHODS AND RESULTS: Through measurements of membrane potential and arterial tension, we show that 11,12-EET induced membrane hyperpolarization and vascular relaxation in endothelium-denuded porcine coronary arteries. These responses were suppressed by S-nitroso-N-acetylpenicillamine (SNAP) and 8-Br-cGMP, an NO donor and a membrane-permeant analogue of cGMP, respectively. The inhibitory actions of SNAP and 8-Br-cGMP on 11,12-EET-induced membrane hyperpolarization and vascular relaxation were reversed by hydroxocobalamin, an NO scavenger; ODQ, a guanylyl cyclase inhibitor; and KT5823, a protein kinase G (PKG) inhibitor. The inhibitory actions of SNAP and 8-bromo cyclic GMP (8-Br-cGMP) on the EET responses were also abrogated by shielding TRPC1-PKG phosphorylation sites with an excessive supply of exogenous PKG substrates, TAT-TRPC1(S172) and TAT-TRPC1(T313). Furthermore, a phosphorylation assay demonstrated that PKG could directly phosphorylate TRPC1 at Ser(172) and Thr(313). In addition, 11,12-EET failed to induce membrane hyperpolarization and vascular relaxation when TRPV4, TRPC1, or KCa1.1 was selectively inhibited. Co-immunoprecipitation studies demonstrated that TRPV4, TRPC1, and KCa1.1 physically associated with each other in smooth muscle cells. CONCLUSION: Our findings demonstrate a novel role of the NO-cGMP-PKG pathway in the inhibition of 11,12-EET-induced smooth muscle hyperpolarization and relaxation via PKG-mediated phosphorylation of TRPC1.
PMID: 25139746External 2231691f894ba696de1310221b0a0dbbb31a7251e75115c265587c3d9d5f507c
Screening Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  Experimental screening Non-experimental screening Reference
TRP channel construct Interactor source
TRP channel Interactor Method Species Region Species Organ/tissue Sample type
TRPV4 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Bkca Inference Prediction 25139746
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
Validation: In vivo validation Toggle 893349bafcc528f8346c51dc3420151d67b0126b2c122dd1017121c03fa0f69b
  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
TRPC1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Bkca Co-immunoprecipitation Porcine coronary artery 25139746
TRPC1 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TRPV4 Co-immunoprecipitation Porcine coronary artery 25139746
TRPV4 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 Bkca Co-immunoprecipitation Porcine coronary artery 25139746
TRPV4 Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4 TRPC1 Co-immunoprecipitation Porcine coronary artery 25139746
(Link 2bd4d11adb659cddf58197a94e201f0a44c55d8d7cb427c624971b42e122c0a4: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

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