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Molecular basis for the inhibition of human NMPRTase, a novel target for anticancer agents

Nature Structural & Molecular Biology volume 13, pages 582–588 (2006)Cite this article

Abstract

Nicotinamide phosphoribosyltransferase (NMPRTase) has a crucial role in the salvage pathway of NAD+ biosynthesis, and a potent inhibitor of NMPRTase, FK866, can reduce cellular NAD+ levels and induce apoptosis in tumors. We have determined the crystal structures at up to 2.1-Å resolution of human and murine NMPRTase, alone and in complex with the reaction product nicotinamide mononucleotide or the inhibitor FK866. The structures suggest that Asp219 is a determinant of substrate specificity of NMPRTase, which is confirmed by our mutagenesis studies. FK866 is bound in a tunnel at the interface of the NMPRTase dimer, and mutations in this binding site can abolish the inhibition by FK866. Contrary to current knowledge, the structures show that FK866 should compete directly with the nicotinamide substrate. Our structural and biochemical studies provide a starting point for the development of new anticancer agents.

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Figure 1: Sequence alignment of NMPRTase with other phosphoribosyltransferases involved in NAD+ biosynthesis.
Figure 2: Structure of human NMPRTase.
Figure 3: Binding mode of NMN and the active site of NMPRTase.
Figure 4: Kinetic characterization of substrate specificity and inhibitor sensitivity of human NMPRTase.
Figure 5: The FK866-binding site of human NMPRTase.

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Acknowledgements

We thank J. Schwanof and R. Abramowitz for setting up the X4A and X4C beamlines at the National Synchrotron Light Source, Y. Shen and S. Xiang for help with data collection at the synchrotron source, M. Matsumoto, J. Buteau and D. Accili for help with the insulin assay, H. Zhang (University of Texas Southwestern Medical Center), M. Wahl (Max-Planck-Institute for Biophysical Chemistry) and S.H. Eom (Gwangju Institute of Science & Technology) for respectively providing the bacterial expression plasmids for human NMN/NAMN adenylyltransferase, E. coli NADS and Helicobacter pylori NADS, and W.W. Cleland for helpful discussions. This work was supported in part by a grant from the US National Institutes of Health.

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  1. Xiao Tao

    Present address: Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, New York, New York, 10021

Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, New York, 10027, New York, USA

    Javed A Khan, Xiao Tao & Liang Tong

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Correspondence to Liang Tong.

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Supplementary information

Supplementary Fig. 1

Structures of NAPRTase and QAPRTase dimers (PDF 760 kb)

Supplementary Fig. 2

Structural changes upon NMN and FK866 binding (PDF 144 kb)

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Khan, J., Tao, X. & Tong, L. Molecular basis for the inhibition of human NMPRTase, a novel target for anticancer agents. Nat Struct Mol Biol 13, 582–588 (2006). https://doi.org/10.1038/nsmb1105

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