ATTENTION: The software behind KU ScholarWorks is being upgraded to a new version. Starting July 15th, users will not be able to log in to the system, add items, nor make any changes until the new version is in place at the end of July. Searching for articles and opening files will continue to work while the system is being updated.
If you have any questions, please contact Marianne Reed at mreed@ku.edu .
Expression, limited proteolysis and preliminary crystallographic analysis of IpaD, a component of the Shigella flexneri type III secretion system
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 |
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 |