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PNAS 101 (45): 15944-15948

Copyright © 2004 by the National Academy of Sciences.

From the Cover


Neuronal nitric oxide synthase negatively regulates xanthine oxidoreductase inhibition of cardiac excitation-contraction coupling

Shakil A. Khan * {dagger}, Kwangho Lee {dagger}, {ddagger}, Khalid M. Minhas * {dagger}, Daniel R. Gonzalez *, Shubha V. Y. Raju *, Ankit D. Tejani *, Dechun Li {ddagger}, Dan E. Berkowitz {ddagger}, and Joshua M. Hare *, §

Departments of *Medicine (Cardiology Division) and {ddagger}Anesthesia and Critical Care Medicine, The Johns Hopkins Medical Institutions, Baltimore, MD 21287

Edited by Solomon H. Snyder, The Johns Hopkins University School of Medicine, Baltimore, MD, and approved September 9, 2004

Received for publication June 10, 2004.

Abstract: Although interactions between superoxide () and nitric oxide underlie many physiologic and pathophysiologic processes, regulation of this crosstalk at the enzymatic level is poorly understood. Here, we demonstrate that xanthine oxidoreductase (XOR), a prototypic superoxide -producing enzyme, and neuronal nitric oxide synthase (NOS1) coimmunoprecipitate and colocalize in the sarcoplasmic reticulum of cardiac myocytes. Deficiency of NOS1 (but not endothelial NOS, NOS3) leads to profound increases in XOR-mediated production, which in turn depresses myocardial excitation–contraction coupling in a manner reversible by XOR inhibition with allopurinol. These data demonstrate a unique interaction between a nitric oxide and an -generating enzyme that accounts for crosstalk between these signaling pathways; these findings demonstrate a direct antioxidant mechanism for NOS1 and have pathophysiologic implications for the growing number of disease states in which increased XOR activity plays a role.

Author contributions: S.A.K., K.L., K.M.M., D.E.B., and J.M.H. designed research; S.A.K., K.L., K.M.M., D.R.G., S.V.Y.R., A.D.T., and D.L. performed research; D.L. and J.M.H. contributed new reagents/analytic tools; S.A.K., K.L., K.M.M., D.R.G., S.V.Y.R., and D.L. analyzed data; and S.A.K., D.E.B., and J.M.H. wrote the paper.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: XOR, xanthine oxidoreductase; NOS, nitric oxide synthase; NOS1, neuronal NOS; NOS3, endothelial NOS; SR, sarcoplasmic reticulum; SERCA, SR Ca2+ ATPase; SL, sarcomere length; DHE, dihydroethidium; DAF, diaminofluorescein; OS, oxidative stress; [Ca2+]i, Ca2+ transient.

{dagger} S.A.K., K.L., and K.M.M. contributed equally to this work.

§ To whom correspondence should be addressed. E-mail: jhare{at}

© 2004 by The National Academy of Sciences of the USA

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