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Allosteric inhibition via R-state destabilization in ATP sulfurylase from Penicillium chrysogenum

Nature Structural Biology volume 9, pages 945–949 (2002)Cite this article

Abstract

The structure of the cooperative hexameric enzyme ATP sulfurylase from Penicillium chrysogenum bound to its allosteric inhibitor, 3′-phosphoadenosine-5′-phosphosulfate (PAPS), was determined to 2.6 Å resolution. This structure represents the low substrate-affinity T-state conformation of the enzyme. Comparison with the high substrate-affinity R-state structure reveals that a large rotational rearrangement of domains occurs as a result of the R-to-T transition. The rearrangement is accompanied by the 17 Å movement of a 10-residue loop out of the active site region, resulting in an open, product release-like structure of the catalytic domain. Binding of PAPS is proposed to induce the allosteric transition by destabilizing an R-state-specific salt linkage between Asp 111 in an N-terminal domain of one subunit and Arg 515 in the allosteric domain of a trans-triad subunit. Disrupting this salt linkage by site-directed mutagenesis induces cooperative inhibition behavior in the absence of an allosteric effector, confirming the role of these two residues.

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Figure 1: Hexameric arrangement of ATP sulfurylase subunits in the R- and T-states.
Figure 2: Conformational changes observed in the R-to-T allosteric transition.
Figure 3: Allosteric inhibitor-binding site.
Figure 4: Velocity versus substrate concentration of the R515A mutant.

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Acknowledgements

The research described in this report was supported by an NSF Research Grant to I.H.S. and A.J.F. and by facilities of the W.M. Keck Foundation Center for Structural Biology at the University of California, Davis. Some of the work reported here was performed at SSRL, which is operated by the Department of Energy, Office of Basic Energy Sciences. The SSRL Biotechnology Program is supported by the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program, and by the Department of Energy, Office of Biological and Environmental Research.

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  1. Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, 95616, California, USA

    Ian J. MacRae, Irwin H. Segel & Andrew J. Fisher

  2. Department of Chemistry, University of California, One Shields Avenue, Davis, 95616, California, USA

    Andrew J. Fisher

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  1. Ian J. MacRae
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Correspondence to Andrew J. Fisher.

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MacRae, I., Segel, I. & Fisher, A. Allosteric inhibition via R-state destabilization in ATP sulfurylase from Penicillium chrysogenum. Nat Struct Mol Biol 9, 945–949 (2002). https://doi.org/10.1038/nsb868

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