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Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease

Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease
Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease
Endothelial dysfunction is associated with decreased NO bioavailability and impaired activation of the NO receptor soluble guanylate cyclase (sGC) in the vasculature and in platelets. Red blood cells (RBCs) are known to produce NO under hypoxic and normoxic conditions; however evidence of expression and/or activity of sGC and downstream signaling pathway including phopshodiesterase (PDE)-5 and protein kinase G (PKG) in RBCs is still controversial. In the present study, we aimed to investigate whether RBCs carry a functional sGC signaling pathway and to address whether this pathway is compromised in coronary artery disease (CAD). Using two independent chromatographic procedures, we here demonstrate that human and murine RBCs carry a catalytically active α1β1-sGC (isoform 1), which converts 32P-GTP into 32P-cGMP, as well as PDE5 and PKG. Specific sGC stimulation by NO+BAY 41-2272 increases intracellular cGMP-levels up to 1000-fold with concomitant activation of the canonical PKG/VASP-signaling pathway. This response to NO is blunted in α1-sGC knockout (KO) RBCs, but fully preserved in α2-sGC KO. In patients with stable CAD and endothelial dysfunction red cell eNOS expression is decreased as compared to aged-matched controls; by contrast, red cell sGC expression/activity and responsiveness to NO are fully preserved, although sGC oxidation is increased in both groups. Collectively, our data demonstrate that an intact sGC/PDE5/PKG-dependent signaling pathway exists in RBCs, which remains fully responsive to NO and sGC stimulators/activators in patients with endothelial dysfunction. Targeting this pathway may be helpful in diseases with NO deficiency in the microcirculation like sickle cell anemia, pulmonary hypertension, and heart failure.
cGMP, nitric oxide, protein kinase G, signaling, non-canonical functions of RBCs
2213-2317
328-337
Cortese-Krott, Miriam M.
7dc9b44c-847c-4196-8866-a3cc0c1dc357
Mergia, Evanthia
5995104b-0e4d-4cf6-9eee-350403c139bb
Kramer, Christian M.
36b6e166-bce9-462a-b000-e36da2e91ee3
Luckstadt, Wiebke
8df7d36d-a8a1-4bb9-8484-81abb11474eb
Yang, Jiangning
c835007c-55fe-4416-a46d-0c3e3a318e6b
Wolff, George
737466da-7d86-4f64-8649-824ce1f88383
Panknin, Christina
447c0654-7fa5-4d4f-9906-5068243b1312
Bracht, Thilo
d7ed9642-8ebf-4034-8fc9-ffc005f228de
Sitek, Barbara
86c165cd-6e4d-4acf-89d5-2e0799526f3f
Pernow, John
5a802c4f-e20b-4661-bb74-6cb16258b76d
Stasch, Johannes-Peter
c64f1e3c-b0cb-4c17-9300-4bd98daa9643
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Koesling, Doris
97899ac0-5ac8-405f-a171-320bd521c656
Kelm, Malte
db2bb062-32d7-4b50-9f65-8ba89ffa5f42
Cortese-Krott, Miriam M., Mergia, Evanthia, Kramer, Christian M., Luckstadt, Wiebke, Yang, Jiangning, Wolff, George, Panknin, Christina, Bracht, Thilo, Sitek, Barbara, Pernow, John, Stasch, Johannes-Peter, Feelisch, Martin, Koesling, Doris and Kelm, Malte (2018) Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease Redox Biology, 14, pp. 328-337. (doi:10.1016/j.redox.2017.08.020).

Cortese-Krott, Miriam M., Mergia, Evanthia, Kramer, Christian M., Luckstadt, Wiebke, Yang, Jiangning, Wolff, George, Panknin, Christina, Bracht, Thilo, Sitek, Barbara, Pernow, John, Stasch, Johannes-Peter, Feelisch, Martin, Koesling, Doris and Kelm, Malte (2018) Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease Redox Biology, 14, pp. 328-337. (doi:10.1016/j.redox.2017.08.020).

Record type: Article

Abstract

Endothelial dysfunction is associated with decreased NO bioavailability and impaired activation of the NO receptor soluble guanylate cyclase (sGC) in the vasculature and in platelets. Red blood cells (RBCs) are known to produce NO under hypoxic and normoxic conditions; however evidence of expression and/or activity of sGC and downstream signaling pathway including phopshodiesterase (PDE)-5 and protein kinase G (PKG) in RBCs is still controversial. In the present study, we aimed to investigate whether RBCs carry a functional sGC signaling pathway and to address whether this pathway is compromised in coronary artery disease (CAD). Using two independent chromatographic procedures, we here demonstrate that human and murine RBCs carry a catalytically active α1β1-sGC (isoform 1), which converts 32P-GTP into 32P-cGMP, as well as PDE5 and PKG. Specific sGC stimulation by NO+BAY 41-2272 increases intracellular cGMP-levels up to 1000-fold with concomitant activation of the canonical PKG/VASP-signaling pathway. This response to NO is blunted in α1-sGC knockout (KO) RBCs, but fully preserved in α2-sGC KO. In patients with stable CAD and endothelial dysfunction red cell eNOS expression is decreased as compared to aged-matched controls; by contrast, red cell sGC expression/activity and responsiveness to NO are fully preserved, although sGC oxidation is increased in both groups. Collectively, our data demonstrate that an intact sGC/PDE5/PKG-dependent signaling pathway exists in RBCs, which remains fully responsive to NO and sGC stimulators/activators in patients with endothelial dysfunction. Targeting this pathway may be helpful in diseases with NO deficiency in the microcirculation like sickle cell anemia, pulmonary hypertension, and heart failure.

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Accepted/In Press date: 17 August 2017
e-pub ahead of print date: 8 September 2017
Published date: April 2018
Keywords: cGMP, nitric oxide, protein kinase G, signaling, non-canonical functions of RBCs

Identifiers

Local EPrints ID: 415003
URI: http://eprints.soton.ac.uk/id/eprint/415003
ISSN: 2213-2317
PURE UUID: 98e5e39e-6953-49f2-86be-5bb251dbd1c4
ORCID for Martin Feelisch: ORCID iD orcid.org/0000-0003-2320-1158

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Date deposited: 20 Oct 2017 16:31
Last modified: 20 Oct 2017 16:31

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Contributors

Author: Miriam M. Cortese-Krott
Author: Evanthia Mergia
Author: Christian M. Kramer
Author: Wiebke Luckstadt
Author: Jiangning Yang
Author: George Wolff
Author: Christina Panknin
Author: Thilo Bracht
Author: Barbara Sitek
Author: John Pernow
Author: Johannes-Peter Stasch
Author: Martin Feelisch ORCID iD
Author: Doris Koesling
Author: Malte Kelm

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