Abstract
The structure of the neuronal nitric oxide synthase inhibitory protein, PIN (protein inhibitor of nNOS), has been determined by NMR spectroscopy. Two N-terminal antiparallel α-helices pack against a four-stranded antiparallel β-sheet in the C-terminal region of the protein, forming a two-layer α/β plait. The three dimensional structure of PIN resembles the fold of the B-chain of aspartylglucosaminidase. A non-prolyl cis peptide bond was found between Pro 52 and Thr 53 of the protein. PIN has a large solvent-exposed hydrophobic surface that contains a cavity and is rimmed with positive charges. This surface may serve as the primary target-binding region for this multi-functional regulatory protein.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Brenman, J.E., & Bredt, D.S. Meth. Enz.. 269, 119–129 (1996).
Rand, M.J., & Li, C.G. Ann. Rev. Physiol. 57, 659–682 (1995).
Garthwaite, J. & Boulton, C.L. Ann. Rev. Physiol. 57, 683–706 (1995).
Lipton, S.A., & Rosenberg, P.A. N. England J. Med. 330, 613–22 (1994).
Choi, D.W. Prog. Brain Res. 100, 47–51 (1994).
Huang, P.L., Dawson, T.M., Bredt, D.S., Snyder, S.H., & Fishman, M.C. Cell 75, 1273–1286 (1993).
Jaffrey, S.R., & Snyder, S.H. Science 274, 774–777 (1996).
Dick, T., Ray, K., Salz, H.K., & Chia, W. Mol. Cell. Biol. 16, 1966–1977 ( 1996).
Greenwood, M.T., Guo, Y., Kumar, U., Beauséjours, S., & Hussain, S.N.A. Biochem. Biophys. Res. Commun. 238, 617–621 (1997).
King, S.M., & Patel-King, R. S. J. Biol. Chem. 270, 11445–11452 (1995).
King, S.M. et al. J. Biol. Chem. 271, 19358– 19366 (1996).
Wüthrich, K. NMR of Proteins and Nucleic Acids. (Wiley, New York; 1986).
Wishart, D.S., & Sykes, B.D. Meth. Enz.. 239, 363–392 (1994).
Holm, L. & Sander, C. J. Mol. Biol. 233, 123–38 (1993).
Oinonen, C., Tikkanen, R., Rouvinen, J., & Peltonen, L. Nature Struct. Biol. 2, 1102–1108 (1995).
Stewart, D.E., Sarkar, A., & Wampler, J.E. J. Mol. Biol. 214, 253– 260 (1990).
Weiss, M.S., Jabs, H, & Hilgenfeld, R. Nature Struct. Biol. 5,– 676 (1998).
Herzberg, O., & Moult, J. Proteins: Struct. Funct. Genet. 11, 223–229 (1991).
McGaughey, G.B., Gagné, M., & Rappé, A.K. J. Biol. Chem. 273, 15458–15463 (1998).
Babu, Y.S., Bugg, C.E., & Cook, W.J. J. Mol. Biol. 204, 191– 204 (1988).
Zhang, M. & Yuan, T. Biochem. Cell Biol. in the press.
Bax, A., & Grzesiek, S. Acc. Chem. Res. 26, 131–138 (1993).
Neri, D., Szyperski, T. Otting, G., Senn, H., & Wüthrich, K. Biochemistry 28, 7510– 7516 (1989).
Kay, L. E., & Bax, A. J. Magn. Res. 86, 110–126 (1990).
Delaglio, F. et al. J. Biomol. NMR 6, 277– 293 (1995).
Garrett, D.S., Powers, R., Gronenborn, A.M., & Clore, G.M. J. Magn. Res. 95, 214–220 (1991).
Nilges, M., Gronenborn, A.M., Brüger, A.T., & Clore, G.M. Prot. Engng. 2, 27–38 ( 1988).
Brünger, A. T. X-PLOR Version 3.1. A system for X-ray crystallography and NMR (Yale University Press, New Haven, Connecticut; 1992).
Kraulis, P.J. J. Appl. Crystallogr. 24, 946–950 (1991).
Nicholls, A. GRASP: graphical representation and analysis of surface properties (Columbia University, New York; 1992).
Laskwoski, R.A., MacArthur, M.W., Moss, D.S. & Thornton, J.M. J. Appl. Crystallogr. 26, 283–291 (1993).
Acknowledgements
We thank L.E. Kay for providing NMR pulse sequences, M. Nilges for the advice in the structure calculation, and D. Smith for careful reading and critical comments of the manuscript. This work is partially supported by an RGC grant to M.Z. from the Research Grant Committee of Hong Kong. The NMR spectrometer used in this work was purchased by the Biotechnology Research Institute, the Hong Kong University of Science and Technology.Hidehito Tochio, Shinya Ohki, Qiang Zhang, Ming Li and Mingjie Zhang
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tochio, H., Ohki, S., Zhang, Q. et al. Solution structure of a protein inhibitor of neuronal nitric oxide synthase . Nat Struct Mol Biol 5, 965–969 (1998). https://doi.org/10.1038/2940
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/2940
This article is cited by
-
Structure-function-folding relationships and native energy landscape of dynein light chain protein: nuclear magnetic resonance insights
Journal of Biosciences (2009)
-
Nitric oxide and the immune response
Nature Immunology (2001)
-
Cystic fibrosis problem probed by proteolysis
Nature Structural Biology (1998)