Expression, purification, crystallization and preliminary crystallographic analysis of MxiH, a subunit of the Shigella flexneri type III secretion system needle
Deane, Janet E.
Cordes, Frank S.
Picking, William D.
Picking, Wendy Lynn
Lea, Susan M.
Blocker, Ariel J.
International Union of Crystallography
Scholarly/refereed, publisher version
Copyright © International Union of Crystallography 2006. This is an open-access article distributed under the terms described at http://journals.iucr.org/services/termsofuse.html.
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A monodisperse truncation mutant of MxiH, the subunit of the needle from the Shigella flexneri type III secretion system (TTSS), has been overexpressed and purified. Crystals were grown of native and selenomethionine-labelled MxiHCΔ5 and diffraction data were collected to 1.9 Å resolution. The crystals belong to space group C2, with unit-cell parameters a = 183.4, b = 28.1, c = 27.8 Å, β = 96.5°. An anomalous difference Patterson map calculated with the data from the SeMet-labelled crystals revealed a single peak on the Harker section v = 0. Inspection of a uranyl derivative also revealed one peak in the isomorphous difference Patterson map on the Harker section v = 0. Analysis of the self-rotation function indicates the presence of a twofold non-crystallographic symmetry axis approximately along a. The calculated Matthews coefficient is 1.9 Å3 Da−1 for two molecules per asymmetric unit, corresponding to a solvent content of 33%.
Deane, J. E., Cordes, F. S., Roversi, P., Johnson, S., Kenjale, R., Picking, W. D., … Blocker, A. (2006). Expression, purification, crystallization and preliminary crystallographic analysis of MxiH, a subunit of the Shigella flexneri type III secretion system needle. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 62(Pt 3), 302–305. http://doi.org/10.1107/S1744309106006555
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