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Crystal structure of a bifunctional transformylase and cyclohydrolase enzyme in purine biosynthesis

Nature Structural Biology volume 8, pages 402–406 (2001)Cite this article

Abstract

ATIC, the product of the purH gene, is a 64 kDa bifunctional enzyme that possesses the final two activities in de novo purine biosynthesis, AICAR transformylase and IMP cyclohydrolase. The crystal structure of avian ATIC has been determined to 1.75 Å resolution by the MAD method using a Se-methionine modified enzyme. ATIC forms an intertwined dimer with an extensive interface of ∼5,000 Å2 per monomer. Each monomer is composed of two novel, separate functional domains. The N-terminal domain (up to residue 199) is responsible for the IMPCH activity, whereas the AICAR Tfase activity resides in the C-terminal domain (200–593). The active sites of the IMPCH and AICAR Tfase domains are ∼50 Å apart, with no structural evidence of a tunnel connecting the two active sites. The crystal structure of ATIC provides a framework to probe both catalytic mechanisms and to design specific inhibitors for use in cancer chemotherapy and inflammation.

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Figure 1: Structure and function of ATIC.
Figure 2: Structure of the IMPCH domain.
Figure 3: The AICAR Tfase domain.
Figure 4: The potassium ion binding site.

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Acknowledgements

This work was supported in part by NIH Grants to I.A.W., S.J.B. and G.P.B. We thank X. Dai and A. Heine for help with data collection and advice during structure determination, M. Rudolph and R. Stanfield for valuable advice and assistance in computational analysis, J. Vergis and K. Bulock for helpful discussions, and the ALS staff of beamline 5.0.2 for guidance during data collection. This is publication 13714-MB from The Scripps Research Institute.

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Authors and Affiliations

  1. Department of Molecular Biology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, 92037, California, USA

    Samantha E. Greasley, Patricia Horton & Ian A. Wilson

  2. Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, 16802, USA

    Joseph Ramcharan & Stephen J. Benkovic

  3. Departments of Pediatrics and Pharmacology, Yale University School of Medicine, 3094 LMP, 333 Cedar Street, P.O. Box 208064, New Haven, 06520-8064, Connecticut, USA

    G. Peter Beardsley

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  1. Samantha E. Greasley
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Correspondence to Ian A. Wilson.

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Greasley, S., Horton, P., Ramcharan, J. et al. Crystal structure of a bifunctional transformylase and cyclohydrolase enzyme in purine biosynthesis. Nat Struct Mol Biol 8, 402–406 (2001). https://doi.org/10.1038/87555

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