1. Molecular Cardiology, Department of Internal Medicine IV, and
2. Institute of Cardiovascular Physiology, University of Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany

Correspondence to: Andreas M. Zeiher¹ Correspondence and requests for materials should be addressed to A.M.Z. (e-mail: Email: Zeiher@em.uni-frankfurt.de).

Abstract

Nitric oxide (NO) produced by the endothelial NO synthase (eNOS) is a fundamental determinant of cardiovascular homesotasis: it regulates systemic blood pressure, vascular remodelling and angiogenesis^{1, 2, 3}. Physiologically, the most important stimulus for the continuous formation of NO is the viscous drag (shear stress) generated by the streaming blood on the endothelial layer^{4, 5, 6, 7, 8}. Although shear-stress-mediated phosphorylation of eNOS is thought to regulate enzyme activity⁹,¹⁰, the mechanism of activation of eNOS is not yet known. Here we demonstrate that the serine/threonine protein kinase Akt/PKB^{11, 12, 13} mediates the activation of eNOS, leading to increased NO production. Inhibition of the phosphatidylinositol-3-OH kinase/Akt pathway or mutation of the Akt site on eNOS protein (at serine 1177) attenuates the serine phosphorylation and prevents the activation of eNOS. Mimicking the phosphorylation of Ser 1177 directly enhances enzyme activity and alters the sensitivity of the enzyme to Ca²⁺, rendering its activity maximal at sub-physiological concentrations of Ca²⁺. Thus, phosphorylation of eNOS by Akt represents a novel Ca²⁺-independent regulatory mechanism for activation of eNOS.