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Research article2003Peer reviewed

Overexpression, purification, crystallization and preliminary X-ray diffraction analysis of the F1 antigen Caf1M-Caf1 chaperone-subunit pre-assembly complex from Yersinia pestis

Zavialov A, Berglund J, Knight SD

Abstract

The F1 capsular antigen of the plague-causing pathogen Yersinia pestis is assembled from monomeric Caf1 subunits via the Caf1M/Caf1A chaperone/usher system. Y. pestis Caf1M-Caf1 chaperone-subunit complex was purified from the periplasm of Escherichia coli cells overexpressing Caf1M and Caf1 and was crystallized in PEG 4000 solution using hanging-drop vapour diffusion. The crystals diffract to a minimum Bragg spacing of 1.8 Angstrom and belong to space group P2(1), with unit-cell parameters a = 36.0, b = 69.2, c = 69.1 Angstrom, beta = 93.0degrees. SeMet-labelled Caf1M-Caf1 complexes were purified and crystallized under the same conditions. The SeMet crystals were identical to the native crystals and diffracted to 1.9 Angstrom. Heavy-atom derivative crystals were prepared by soaking in 10 mM K2PtCl4, giving two Pt sites per complex. The experimental electron-density map was obtained by a combination of MAD and MIR methods using both Se- and Pt-derivative crystals

Published in

Acta Crystallographica Section D: Biological Crystallography
2003, Volume: 59, pages: 359-362
Publisher: BLACKWELL MUNKSGAARD

      SLU Authors

    • Zavialov, Anton

      • Department of Molecular Biology, Swedish University of Agricultural Sciences

      • Berglund, Jenny

        • Department of Molecular Biology, Swedish University of Agricultural Sciences

        • Knight, Stefan David

          • Department of Molecular Biology, Swedish University of Agricultural Sciences

        UKÄ Subject classification

        Food Science

        Publication identifier

        DOI: https://doi.org/10.1107/S0907444902021054

        Permanent link to this page (URI)

        https://res.slu.se/id/publ/760