dc.contributor.author | Johnson, Steven | |
dc.contributor.author | Roversi, Pietro | |
dc.contributor.author | Espina, Marianela | |
dc.contributor.author | Deane, Janet E. | |
dc.contributor.author | Birket, Susan | |
dc.contributor.author | Picking, William D. | |
dc.contributor.author | Picking, Wendy Lynn | |
dc.contributor.author | Lea, Susan M. | |
dc.date.accessioned | 2016-07-21T18:07:05Z | |
dc.date.available | 2016-07-21T18:07:05Z | |
dc.date.issued | 2006-08-11 | |
dc.identifier.citation | Johnson, 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. http://doi.org/10.1107/S1744309106027047 | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/21169 | |
dc.description.abstract | aD, 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.publisher | International Union of Crystallography | en_US |
dc.rights | 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. | en_US |
dc.subject | IpaD | en_US |
dc.subject | Type III secretion | en_US |
dc.subject | Shigella flexneri | en_US |
dc.title | Expression, limited proteolysis and preliminary crystallographic analysis of IpaD, a component of the Shigella flexneri type III secretion system | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Picking, Wendy Lynn | |
kusw.kuauthor | Espina, Marianela | |
kusw.kuauthor | Birket, Susan | |
kusw.kuauthor | Picking, William D. | |
kusw.kudepartment | Pharmaceutical Chemistry | en_US |
kusw.oanotes | Per 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
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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
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Published source must be acknowledged with citation | en_US |
dc.identifier.doi | 10.1107/S1744309106027047 | en_US |
dc.identifier.orcid | https://orcid.org/0000-0001-7998-0643 | |
kusw.oaversion | Scholarly/refereed, publisher version | en_US |
kusw.oapolicy | This item does not meet KU Open Access policy criteria. | en_US |
dc.rights.accessrights | openAccess | |