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Substrate-assisted catalysis in cytochrome P450eryF

Nature Structural Biology volume 3, pages 632–637 (1996)Cite this article

Abstract

A highly conserved threonine in the active site of cytochromes P450 has been proposed to participate in O2 binding and cleavage. Cytochrome P450eryF is unusual in having alanine in place of this threonine and an ordered active site water molecule (Wat 519) which is hydrogen bonded to the substrate 5-hydroxyl group and is in position to operate as an acid catalyst required for cleaving dioxygen. To asses the role of this alanine residue and Wat 519 in catalysis, two mutant forms of P450eryF (Ala→Ser, Ala→Thr) and a substrate analogue lacking a 5-hydroxyl group were examined using kinetic, spectral and crystallographic techniques. In each case decreased catalytic activity was correlated with a loss or repositioning of Wat 519. These findings suggest that P450eryF utilizes the substrate to assist in the acid-catalysed dioxygen bond cleavage reaction.

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry and Department of Physiology and Biophysics, University of California, Irvine, California, 92717, USA

    Jill R. Cupp-Vickery & Thomas L. Poulos

  2. School of Pharmacy and Department of Bacteriology, University of Wisconsin, Madison, Wisconsin, 53706, USA

    Oksoo Han & C. Richard Hutchinson

Authors
  1. Jill R. Cupp-Vickery
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  2. Oksoo Han
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  3. C. Richard Hutchinson
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  4. Thomas L. Poulos
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Cupp-Vickery, J., Han, O., Hutchinson, C. et al. Substrate-assisted catalysis in cytochrome P450eryF. Nat Struct Mol Biol 3, 632–637 (1996). https://doi.org/10.1038/nsb0796-632

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