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Structure-based design of a lysozyme with altered catalytic activity

Nature Structural Biology volume 2, pages 1007–1011 (1995)Cite this article

Abstract

Here we show that the substitution Thr 26→His in the active site of T4 lysozyme causes the product to change from the α- to the β-anomer. This implies an alteration in the catalytic mechanism of the enzyme. From the change in product, together with inspection of relevant crystal structures, it is inferred that wild-type T4 lysozyme is an anomer-inverting enzyme with a single displacement mechanism in which water attacks from the α-side of the substrate. In contrast, the mutant T26H is an anomer-retaining enzyme with an apparently double displacement mechanism in which a water molecule attacks from the opposite side of the substrate. The results also show that the mechanism of wild-type T4 lysozyme differs from that of hen egg-white lysozyme even though both enzymes are presumed to have evolved from a common precursor.

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Author notes

  1. Ryota Kuroki

    Present address: Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., 1-13-5, Fukuura, Kanazawa-ku Yokohama, 236, Japan

Authors and Affiliations

  1. Institute of Molecular Biology, Howard Hughes Medical Institute and Department of Physics, University of Oregon, Eugene, Oregon, 97403, USA

    Ryota Kuroki, Larry H. Weaver & Brian W. Matthews

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  2. Larry H. Weaver
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  3. Brian W. Matthews
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Kuroki, R., Weaver, L. & Matthews, B. Structure-based design of a lysozyme with altered catalytic activity. Nat Struct Mol Biol 2, 1007–1011 (1995). https://doi.org/10.1038/nsb1195-1007

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