Crystallization of the cytotoxic domain of a ribosome-inactivating colicin in complex with its immunity protein

S. Carr, D. Walker, R. James, C. Kleanthous, A. M. Hemmings

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    Abstract

    The complex between the ribonuclease domain of the ribosome-inactivating colicin E3 and its protein inhibitor, the cognate immunity Im3, has been crystallized and preliminary X-ray characterization has been performed. Single crystals of the 1:1 complex were grown from hanging-drop vapour-diffusion experiments using 2-propanol as a precipitant. The space group is P3121 or P3221, with unit-cell parameters a = b = 93.7, c = 76.2 Å. When cryocooled, these crystals diffract to a resolution of 2.4 Å. A search for suitable conventional heavy-atom derivatives was unsuccessful and so Im3 mutants containing engineered cysteine or methionine residues have been produced for mercury soaks and selenomethionine-labelling experiments, respectively.

    Original languageEnglish
    Pages (from-to)1630-1633
    Number of pages4
    JournalActa Crystallographica Section D: Biological Crystallography
    Volume56
    Issue number12
    DOIs
    Publication statusPublished - 2000

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