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Solution structure of a protein inhibitor of neuronal nitric oxide synthase

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.

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Figure 1: Sequence alignment of PIN and its homologs from various species.
Figure 2: Stereo view showing the best-fit superposition of the backbone atoms (N, Cα, and C') of the final 20 structures of PIN.
Figure 3: Portion of the 13C-separated 3D NOESY spectrum of 15N,13C-labelled PIN showing the NOE crosspeaks of the αH of Pro 52 with the αH and the βH of Thr 53 respectively.
Figure 4: Molecular surface representation of PIN showing a, the solvent-exposed hydrophobic surface area and b, the hydrophilic surface, rotated by 180o with respect to the view in (a).

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References

  1. Brenman, J.E., & Bredt, D.S. Meth. Enz.. 269, 119–129 (1996).

    Article  CAS  Google Scholar 

  2. Rand, M.J., & Li, C.G. Ann. Rev. Physiol. 57, 659–682 (1995).

    Article  CAS  Google Scholar 

  3. Garthwaite, J. & Boulton, C.L. Ann. Rev. Physiol. 57, 683–706 (1995).

    Article  CAS  Google Scholar 

  4. Lipton, S.A., & Rosenberg, P.A. N. England J. Med. 330, 613–22 (1994).

    Article  CAS  Google Scholar 

  5. Choi, D.W. Prog. Brain Res. 100, 47–51 (1994).

    Article  CAS  Google Scholar 

  6. Huang, P.L., Dawson, T.M., Bredt, D.S., Snyder, S.H., & Fishman, M.C. Cell 75, 1273–1286 (1993).

    Article  CAS  Google Scholar 

  7. Jaffrey, S.R., & Snyder, S.H. Science 274, 774–777 (1996).

    Article  CAS  Google Scholar 

  8. Dick, T., Ray, K., Salz, H.K., & Chia, W. Mol. Cell. Biol. 16, 1966–1977 ( 1996).

    Article  CAS  Google Scholar 

  9. Greenwood, M.T., Guo, Y., Kumar, U., Beauséjours, S., & Hussain, S.N.A. Biochem. Biophys. Res. Commun. 238, 617–621 (1997).

    Article  CAS  Google Scholar 

  10. King, S.M., & Patel-King, R. S. J. Biol. Chem. 270, 11445–11452 (1995).

    Article  CAS  Google Scholar 

  11. King, S.M. et al. J. Biol. Chem. 271, 19358– 19366 (1996).

    Article  CAS  Google Scholar 

  12. Wüthrich, K. NMR of Proteins and Nucleic Acids. (Wiley, New York; 1986).

    Book  Google Scholar 

  13. Wishart, D.S., & Sykes, B.D. Meth. Enz.. 239, 363–392 (1994).

    Article  CAS  Google Scholar 

  14. Holm, L. & Sander, C. J. Mol. Biol. 233, 123–38 (1993).

    Article  CAS  Google Scholar 

  15. Oinonen, C., Tikkanen, R., Rouvinen, J., & Peltonen, L. Nature Struct. Biol. 2, 1102–1108 (1995).

    Article  CAS  Google Scholar 

  16. Stewart, D.E., Sarkar, A., & Wampler, J.E. J. Mol. Biol. 214, 253– 260 (1990).

    Article  CAS  Google Scholar 

  17. Weiss, M.S., Jabs, H, & Hilgenfeld, R. Nature Struct. Biol. 5,– 676 (1998).

    Article  Google Scholar 

  18. Herzberg, O., & Moult, J. Proteins: Struct. Funct. Genet. 11, 223–229 (1991).

    Article  CAS  Google Scholar 

  19. McGaughey, G.B., Gagné, M., & Rappé, A.K. J. Biol. Chem. 273, 15458–15463 (1998).

    Article  CAS  Google Scholar 

  20. Babu, Y.S., Bugg, C.E., & Cook, W.J. J. Mol. Biol. 204, 191– 204 (1988).

    Article  CAS  Google Scholar 

  21. Zhang, M. & Yuan, T. Biochem. Cell Biol. in the press.

  22. Bax, A., & Grzesiek, S. Acc. Chem. Res. 26, 131–138 (1993).

    Article  CAS  Google Scholar 

  23. Neri, D., Szyperski, T. Otting, G., Senn, H., & Wüthrich, K. Biochemistry 28, 7510– 7516 (1989).

    Article  CAS  Google Scholar 

  24. Kay, L. E., & Bax, A. J. Magn. Res. 86, 110–126 (1990).

    CAS  Google Scholar 

  25. Delaglio, F. et al. J. Biomol. NMR 6, 277– 293 (1995).

    Article  CAS  Google Scholar 

  26. Garrett, D.S., Powers, R., Gronenborn, A.M., & Clore, G.M. J. Magn. Res. 95, 214–220 (1991).

    CAS  Google Scholar 

  27. Nilges, M., Gronenborn, A.M., Brüger, A.T., & Clore, G.M. Prot. Engng. 2, 27–38 ( 1988).

    Article  CAS  Google Scholar 

  28. Brünger, A. T. X-PLOR Version 3.1. A system for X-ray crystallography and NMR (Yale University Press, New Haven, Connecticut; 1992).

    Google Scholar 

  29. Kraulis, P.J. J. Appl. Crystallogr. 24, 946–950 (1991).

    Article  Google Scholar 

  30. Nicholls, A. GRASP: graphical representation and analysis of surface properties (Columbia University, New York; 1992).

    Google Scholar 

  31. Laskwoski, R.A., MacArthur, M.W., Moss, D.S. & Thornton, J.M. J. Appl. Crystallogr. 26, 283–291 (1993).

    Article  Google Scholar 

Download references

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

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Correspondence to Mingije Zhang.

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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

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