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The Journal of Neuroscience, November 17, 2004, 24(46):10454-10465; doi:10.1523/JNEUROSCI.2265-04.2004

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Cellular/Molecular
Nitric Oxide Synthase (NOS)-Interacting Protein Interacts with Neuronal NOS and Regulates Its Distribution and Activity

Jacqueline Dreyer,^1 * Michael Schleicher,^2 * Anke Tappe,¹ Kirstin Schilling,² Thomas Kuner,³ Grace Kusumawidijaja,¹ Werner Müller-Esterl,² Stefanie Oess,² and Rohini Kuner¹

¹Department of Molecular Pharmacology, Pharmacology Institute, University of Heidelberg, 69120 Heidelberg, Germany, ²Institute for Biochemistry II, University of Frankfurt, 60590 Frankfurt, Germany, and ³Department of Cell Physiology, Max-Planck Institute for Medical Research, 69120 Heidelberg, Germany

Mechanisms governing the activity of neuronal nitric oxide synthase (nNOS), the major source of nitric oxide (NO) in the nervous system, are not completely understood. We report here a protein-protein interaction between nNOS and NOSIP (nitric oxide synthase-interacting protein) in rat brain in vivo. NOSIP and nNOS are concentrated in neuronal synapses and demonstrate significant colocalization in various regions of the central and peripheral nervous systems. NOSIP produces a significant reduction in nNOS activity in a neuroepithelioma cell line stably expressing nNOS. Furthermore, overexpression of NOSIP in cultured primary neurons reduces the availability of nNOS in terminal dendrites. These results thus suggest that the interaction between NOSIP and nNOS is functionally involved in endogenous mechanisms regulating NO synthesis. Furthermore, we found that the subcellular distribution and expression levels of NOSIP are dynamically regulated by neuronal activity in vitro as well as in vivo, suggesting that NOSIP may contribute to a mechanism via which neuronal activity regulates the synaptic availability and activity of nNOS.

Key words: nitric oxide; synaptic spines; hippocampal neurons; protein-protein interactions; seizures; inflammatory pain

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Received June 9, 2004; revised October 7, 2004; accepted October 7, 2004.

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