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Transition-state analogs as inhibitors of human and malarial hypoxanthine-guanine phosphoribosyltransferases

Nature Structural Biology volume 6, pages 582–587 (1999)Cite this article

Abstract

The proposed transition state for hypoxanthine-guanine phosphoribosyltransferases (HGPRTs) has been used to design and synthesize powerful inhibitors that contain features of the transition state. The iminoribitols (1S)-1-(9-deazahypoxanthin-9-yl)-1,4-dideoxy-1, 4-imino-D-ribitol 5-phosphate (immucillinHP) and (1S)-1-(9-deazaguanin-9-yl)-1,4-dideoxy-1, 4-imino-D-ribitol 5-phosphate (immucillinGP) are the most powerful inhibitors yet reported for both human and malarial HGPRTs. Equilibrium binding constants are >1,000-fold tighter than the binding of the nucleotide substrate. The NMR spectrum of malaria HGXPRT in the Michaelis complex reveals downfield hydrogen-bonded protons. The chemical shifts move farther downfield with bound inhibitor. The inhibitors are lead compounds for species-specific antibiotics against parasitic protozoa. The high-resolution crystal structure of human HGPRT with immucillinGP is reported in the companion paper.

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Figure 1: Substrate (IMP), proposed transition state and inhibitors for HGPRT.
Figure 2: a, Estimate of the transition-state binding energy for IMP and PPi at the transition state of human HGPRT.
Figure 3: Inhibition of human HGPRT by immucillin nucleotides.
Figure 4: Release of immucillins from the HGPRT complexes.
Figure 5: Proton NMR spectra of the downfield protons in malarial HGXPRT, the Michaelis complex and the complex with immucillin nucleotides.
Figure 6: Comparison of a, human HGPRT–immucillinGP–MgPPi and b, Trypanosoma cruzi HPRT–formycin-B base–MgPRPP in the crystal structures.

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Acknowledgements

Research grants and training grants from the National Institutes of Health, and the New Zealand Foundation for Research, Science and Technology supported this research. The NMR facility was supported in part by grants from the NSF and the Howard Hughes Medical Institute Biomedical Research Support Program for Medical Schools. We thank A. Sauve for the enzymatic phosphorylation of [7-15N]immucillinHP.

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Authors and Affiliations

  1. Department of Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, 10461, New York, USA

    Caroline M. Li, Gregory Kicska, Mark E. Girvin & Vern L. Schramm

  2. Carbohydrate Chemistry Team, Industrial Research Ltd., Lower Hutt, New Zealand

    Peter C. Tyler & Richard H. Furneaux

  3. Department of Biochemistry and Fels Research Institute, Temple University School of Medicine, Philadelphia, 19140, Pennsylvania, USA

    Yiming Xu & Charles Grubmeyer

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  1. Caroline M. Li
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Correspondence to Vern L. Schramm.

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Li, C., Tyler, P., Furneaux, R. et al. Transition-state analogs as inhibitors of human and malarial hypoxanthine-guanine phosphoribosyltransferases. Nat Struct Mol Biol 6, 582–587 (1999). https://doi.org/10.1038/9367

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