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PDZ domain of neuronal nitric oxide synthase recognizes novel C-terminal peptide sequences

Nature Biotechnology volume 15, pages 336–342 (1997)Cite this article

Abstract

PDZ domains are multifunctional protein-interaction motifs that often bind to the C-terminus of protein targets. Nitric oxide (NO), an endogenous signaling molecule, plays critical roles in nervous, immune, and cardiovascular function. Although there are numerous physiological functions for neuron-derived NO, produced primarily by the neuronal NO synthase (nNOS), excess nNOS activity mediates brain injury in cerebral ischemia and in animal models of Parkinson's disease. Subcellular localization of nNOS activity must therefore be tightly regulated. To determine ligands for the PDZ domain of nNOS, we screened 13 billion distinct peptides and found that the nNOS-PDZ domain binds tightly to peptides ending Asp-X-Val. This differs from the only known (ThiYSer)-X-Val consensus that interacts with PDZ domains from PSD-95. Preference for Asp at the -2 peptide position is mediated by Tyr-77 of nNOS. A Y77D78 to H77E7B substitution changes the binding specificity from Asp-X-Val to Thr-X-Val. Guided by the Asp-X-Val consensus, candidate nNOS interacting proteins have been identified including glutamate and melatonin receptors. Our results demonstrate that PDZ domains have distinct peptide binding specificity.

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Author notes

  1. Min Li: Corresponding author: (e-mail: min_li®qmail.bs.jhu.edu).

Authors and Affiliations

  1. Departments of Physiology and Neuroscience,

    Nicole L. Stricker & Min Li

  2. Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD, 21205

    Douglas E. Bassett Jr.

  3. Department of Physiology and Program in Biomedical Sciences, University of California, San Francisco, CA, 94143

    Karen S. Christopherson, Byungdoo A. Yi & David S. Bredt

  4. Department of Molecular Biology, AffymaxResearch Institute, PaloAlto, CA, 94304

    Peter J. Schatz, Ronald W. Raab & Glenn Dawes

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  1. Nicole L. Stricker
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  2. Karen S. Christopherson
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Stricker, N., Christopherson, K., Yi, B. et al. PDZ domain of neuronal nitric oxide synthase recognizes novel C-terminal peptide sequences. Nat Biotechnol 15, 336–342 (1997). https://doi.org/10.1038/nbt0497-336

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  • DOI: https://doi.org/10.1038/nbt0497-336

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