DINITROBENZENES STIMULATE ELECTRON FLUX WITHIN NEURONAL NITRIC OXIDE SYNTHASE IN THE ABSENCE OF CALMODULIN

Overview

Journal Int J Biomed Res

Date 2014 Jan 24

PMID 24453460

Authors

Chintamani N Joshi

David A Tulis

Richard T Miller

Affiliations

Soon will be listed here.

Abstract

Efficient electron transfer and conversion of -arginine to -citrulline and nitric oxide (NO) by neuronal nitric oxide synthase (nNOS) requires calmodulin (CaM) binding. The present study focused on electron transfer ability of resting state CaM-free nNOS in presence of dinitrobenzene isomers (DNBs). NADPH oxidation (NADPH ) and acetylated cytochrome- reduction (AcCyt- ) catalyzed by nNOS and the CaM binding sequence-deficient nNOS reductase construct (nNOS-FP) were estimates of total electron flux and [Formula: see text] production, respectively. All the DNBs (-, -, -) independently stimulated rates of NADPH by CaM-free nNOS and by nNOS-FP in isomer- and concentration-dependent manner. Blocking nNOS heme by imidazole or -arginine did not affect CaM-free nNOS-catalyzed NADPH stimulated by DNBs. This stimulated electron flux by DNBs did not support NO formation by CaM-free nNOS. The DNBs, like FeCN, extract electrons from both FMN and FAD of the nNOS reductase domain. All three DNBs greatly stimulated nNOS and nNOS-FP catalyzed AcCyt- that was significantly inhibited by SOD demonstrating [Formula: see text] formation. Thus, in presence of DNBs, resting-state CaM-deficient nNOS efficiently transfers electrons generating [Formula: see text], inferring that additional metabolic roles for nNOS exist that are not yet explored.

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