Active site residues in m-calpain: identification by site-directed mutagenesis

J. Simon C. Arthur, Sherry Gauthier, John S. Elce

    Research output: Contribution to journalArticle

    49 Citations (Scopus)

    Abstract

    Site-directed mutagenesis was used to alter putative active site residues in the large subunit of calpain, and the activity of the mutants was measured following coexpression in E. coli of both calpain subunits and purification of the resultant dimers. Mutants Cys105Ser, His262Ala and Asn286Ala had no activity. Together with sequence comparisons among cysteine proteinases, the results suggest that these residues constitute the catalytic triad in calpain. Mutants Asn286Asp and Trp288Tyr had low activity, consistent with interaction of these residues with His262.

    Original languageEnglish
    Pages (from-to)397-400
    Number of pages4
    JournalFEBS Letters
    Volume368
    Issue number3
    DOIs
    Publication statusPublished - 24 Jul 1995

    Keywords

    • Rats
    • Mutagenesis, Site-Directed
    • Animals
    • Calcium
    • Humans
    • Escherichia coli
    • Calpain
    • Cloning, Molecular
    • Binding Sites

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