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dc.contributor.authorJohnson, Steven
dc.contributor.authorRoversi, Pietro
dc.contributor.authorEspina, Marianela
dc.contributor.authorDeane, Janet E.
dc.contributor.authorBirket, Susan
dc.contributor.authorPicking, William D.
dc.contributor.authorPicking, Wendy Lynn
dc.contributor.authorLea, Susan M.
dc.identifier.citationJohnson, S., Roversi, P., Espina, M., Deane, J. E., Birket, S., Picking, W. D., … Lea, S. M. (2006). Expression, limited proteolysis and preliminary crystallographic analysis of IpaD, a component of the Shigella flexneri type III secretion system. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 62(Pt 9), 865–868.
dc.description.abstractaD, the putative needle-tip protein of the Shigella flexneri type III secretion system, has been overexpressed and purified. Crystals were grown of the native protein in space group P212121, with unit-cell parameters a = 55.9, b = 100.7, c = 112.0 Å, and data were collected to 2.9 Å resolution. Analysis of the native Patterson map revealed a peak at 50% of the origin on the Harker section v = 0.5, suggesting twofold non-crystallographic symmetry parallel to the b crystallographic axis. As attempts to derivatize or grow selenomethionine-labelled protein crystals failed, in-drop proteolysis was used to produce new crystal forms. A trace amount of subtilisin Carlsberg was added to IpaD before sparse-matrix screening, resulting in the production of several new crystal forms. This approach produced SeMet-labelled crystals and diffraction data were collected to 3.2 Å resolution. The SeMet crystals belong to space group C2, with unit-cell parameters a = 139.4, b = 45.0, c = 99.5 Å, β = 107.9°. An anomalous difference Patterson map revealed peaks on the Harker section v = 0, while the self-rotation function indicates the presence of a twofold noncrystallographic symmetry axis, which is consistent with two molecules per asymmetric unit.en_US
dc.publisherInternational Union of Crystallographyen_US
dc.rightsCopyright © International Union of Crystallography 2006. This is an open-access article distributed under the terms described at
dc.subjectType III secretionen_US
dc.subjectShigella flexnerien_US
dc.titleExpression, limited proteolysis and preliminary crystallographic analysis of IpaD, a component of the Shigella flexneri type III secretion systemen_US
kusw.kuauthorPicking, Wendy Lynn
kusw.kuauthorEspina, Marianela
kusw.kuauthorBirket, Susan
kusw.kuauthorPicking, William D.
kusw.kudepartmentPharmaceutical Chemistryen_US
kusw.oanotesPer SHERPA/RoMEO 7/21/2016: Author's Pre-print: green tick author can archive pre-print (ie pre-refereeing) Author's Post-print: green tick author can archive post-print (ie final draft post-refereeing) Publisher's Version/PDF: green tick author can archive publisher's version/PDF General Conditions:

On author's personal website, employer's website, employer's repository, or subject-based repository Publisher's version/PDF may be used (authorised electronic re-print) (preferred) Publisher's version/PDF (authorised electronic re-print) on PubMed Central and related servers Pre-print must acknowledge submission to journal Must link to publisher version on IUCr server Published source must be acknowledged with citation
kusw.oaversionScholarly/refereed, publisher versionen_US
kusw.oapolicyThis item does not meet KU Open Access policy criteria.en_US

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