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

Redox Biol. 2018 Apr;14:328-337. doi: 10.1016/j.redox.2017.08.020. Epub 2017 Sep 8.

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.

Keywords: Nitric oxide; Non -canonical functions of RBCs; Protein kinase G; Signaling; cGMP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Animals
  • Coronary Artery Disease / metabolism*
  • Cyclic GMP / metabolism
  • Cyclic GMP-Dependent Protein Kinases / metabolism
  • Erythrocytes / metabolism*
  • Humans
  • Mice
  • Middle Aged
  • Nitric Oxide / metabolism
  • Signal Transduction
  • Soluble Guanylyl Cyclase / analysis
  • Soluble Guanylyl Cyclase / metabolism*

Substances

  • Nitric Oxide
  • Cyclic GMP-Dependent Protein Kinases
  • Soluble Guanylyl Cyclase
  • Cyclic GMP