Nitric oxide can acutely modulate its biosynthesis through a negative feedback mechanism on L-arginine transport in cardiac myocytes

Zhou J, Kim DD, Peluffo RD. Nitric oxide can acutely modulate its biosynthesis through a negative feedback mechanism on L-arginine transport in cardiac myocytes. Am J Physiol Cell Physiol 299: C230-C239, 2010. First published May 26, 2010; doi: 10.1152/ajpcell.00077.2010.Nitric oxide (NO) plays a central role as a cellular signaling molecule in health and disease. In the heart, NO decreases the rate of spontaneous beating and the velocity and extent of shortening and accelerates the velocity of relengthening. Since the cationic amino acid L-arginine (L-Arg) is the substrate for NO production by NO synthases (NOS), we tested whether the transporters that mediate L-Arg import in cardiac muscle cells represent an intervention point in the regulation of NO synthesis. Electrical currents activated by L-Arg with low apparent affinity in whole cell voltage-clamped rat cardiomyocytes were found to be rapidly and reversibly inhibited by NO donors. Radiotracer uptake studies performed on cardiac sarcolemmal vesicles revealed the presence of high-affinity/low-capacity and low-affinity/high-capacity components of cationic amino acid transport that were inhibited by the NO donor S-nitroso-N-acetyl-DL-penicillamine. NO inhibited uptake in a noncompetitive manner with K-i values of 275 and 827 nM for the high- and low-affinity component, respectively. Fluorescence spectroscopy experiments showed that millimolar concentrations of L-Arg initially promoted and then inhibited the release of endogenous NO in cardiomyocytes. Likewise, L-Arg currents measured in cardiac myocytes voltage clamped in the presence of 460 nM free intracellular Ca2+, a condition in which a Ca-CaM complex should activate endogenous NO production, showed fast activation followed by inhibition of L-Arg transport. The NOS inhibitor N-nitro-L-arginine methyl ester, but not blockers of downstream reactions, specifically removed this inhibitory component. These results demonstrate that NO acutely regulates its own biosynthesis by modulating the availability of L-Arg via cationic amino acid transporters.