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    A pulse-radiolysis approach to fast reductive cleavage of a disulfide bond to uncage enzyme activity

    Milanesi, Lilia and Tomas, Salvador and Hunter, C.A. and Weinstein, J.A. and Edge, R. and Navaratnam, S. and Waltho, J.P. and Best, J. (2008) A pulse-radiolysis approach to fast reductive cleavage of a disulfide bond to uncage enzyme activity. Free Radical Biology and Medicine 45 (9), pp. 1271-1278. ISSN 0891-5849.

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    Abstract

    The essential thiol of the enzyme papain has been caged by linking to an aromatic thiol. The resulting caged protein is inactive but enzymatic activity is fully restored upon chemical cleavage of the protective disulfide bond. We have exploited the chemistry of this disulfide bond to uncage papain by pulse radiolysis. We have shown that up to 10% of the enzyme activity can be restored by reductive pulse radiolysis. This approach has been tested on a small-molecule model system, and experiments on this model compound show that pulse radiolysis of the mixed cysteine-aromatic disulfide results in selective reduction of the disulfide bond to generate a thiol in 10–20% yield, consistent with the radiolytically restored activity of the caged papain quantified by the biochemical assay.

    Metadata

    Item Type: Article
    Keyword(s) / Subject(s): Biological activity, free radicals, reductive pulse radiolysis, papain, uncaging
    School: Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences
    Research Centres and Institutes: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Administrator
    Date Deposited: 04 Aug 2010 14:09
    Last Modified: 02 Aug 2023 16:49
    URI: https://eprints.bbk.ac.uk/id/eprint/1136

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