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Solution structure of a protein inhibitor of neuronal nitric oxide synthase

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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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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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  1. Department of Biochemistry, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, P.R. China

    Hidehito Tochio, Shinya Ohki, Qiang Zhang, Ming Li & Mingije Zhang

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  1. Hidehito Tochio
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  2. Shinya Ohki
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  3. Qiang Zhang
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  4. Ming Li
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  5. Mingije 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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