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Conversion of the substrate specificity of mouse proteinase granzyme B

Nature Structural Biology volume 1, pages 364–367 (1994)Cite this article

Abstract

Mouse granzyme B is the prototypic member of a subfamily of serine proteinases expressed in cytolytic lymphocytes. Molecular modelling of granzyme B indicated that the side chain of Arg 208 partially fills the specificity pocket, thus predicting the preference of this enzyme for substrates containing acidic side chains, a feature unique among eukaryotic serine proteinases. Replacement of Arg 208 with glycine results in an enzyme lacking this activity, but which is able to hydrolyze hydrophobic substrates. These results demonstrate unequivocally that the substrate preference of granzyme B is determined by a positive charge in the specificity pocket and also represent one of the few examples of rational and efficient alteration of serine proteinase substrate-specificity following a single amino acid substitution.

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Authors and Affiliations

  1. Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G2H7, Canada

    Antonio Caputo, Michael N.G. James & R. Chris Bleackley

  2. School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, 30332-0400, USA

    James C. Powers

  3. Cell and Molecular Biology Graduate Program and Department of Microbiology School of Medicine, University of Nevada, Reno, Nevada, 89557-0046, USA

    Dorothy Hudig

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  1. Antonio Caputo
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  2. Michael N.G. James
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  4. Dorothy Hudig
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  5. R. Chris Bleackley
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Caputo, A., James, M., Powers, J. et al. Conversion of the substrate specificity of mouse proteinase granzyme B. Nat Struct Mol Biol 1, 364–367 (1994). https://doi.org/10.1038/nsb0694-364

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