Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system.
A construct consisting of residues 10-310 of BipD, a component of the Burkholderia pseudomallei type III secretion system (T3SS), has been overexpressed as a GST fusion, cleaved from the GST tag and purified. Crystals were grown of native and selenomethionine-labelled BipD. The crystals grow in two...
| Váldodahkkit: | , , , , , |
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| Materiálatiipa: | Journal article |
| Giella: | English |
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2006
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| _version_ | 1826259796355973120 |
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| author | Roversi, P Johnson, S Field, T Deane, J Galyov, E Lea, S |
| author_facet | Roversi, P Johnson, S Field, T Deane, J Galyov, E Lea, S |
| author_sort | Roversi, P |
| collection | OXFORD |
| description | A construct consisting of residues 10-310 of BipD, a component of the Burkholderia pseudomallei type III secretion system (T3SS), has been overexpressed as a GST fusion, cleaved from the GST tag and purified. Crystals were grown of native and selenomethionine-labelled BipD. The crystals grow in two different polymorphs from the same condition. The first polymorph belongs to space group C222, with unit-cell parameters a = 103.98, b = 122.79, c = 49.17 A, a calculated Matthews coefficient of 2.4 A(3) Da(-1) (47% solvent content) and one molecule per asymmetric unit. The second polymorph belongs to space group P2(1)2(1)2, with unit-cell parameters a = 136.47, b = 89.84, c = 50.15 A, and a calculated Matthews coefficient of 2.3 A(3) Da(-1) (45% solvent content) for two molecules per asymmetric unit (analysis of the self-rotation function indicates the presence of a weak twofold non-crystallographic symmetry axis in this P2(1)2(1)2 form). The native crystals of both forms give diffraction data to 2.7 A resolution, while the SeMet-labelled P2(1)2(1)2 crystals diffract to 3.3 A resolution. A K(2)PtCl(4) derivative of the P2(1)2(1)2 form was also obtained and data were collected to 2.7 A with radiation of wavelength lambda = 0.933 A. The Pt-derivative anomalous difference Patterson map revealed two self-peaks on the Harker sections. |
| first_indexed | 2024-03-06T18:55:27Z |
| format | Journal article |
| id | oxford-uuid:11acf923-f266-40ad-9068-8dd5ac8fc2f5 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:55:27Z |
| publishDate | 2006 |
| record_format | dspace |
| spelling | oxford-uuid:11acf923-f266-40ad-9068-8dd5ac8fc2f52022-03-26T10:03:34ZExpression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:11acf923-f266-40ad-9068-8dd5ac8fc2f5EnglishSymplectic Elements at Oxford2006Roversi, PJohnson, SField, TDeane, JGalyov, ELea, SA construct consisting of residues 10-310 of BipD, a component of the Burkholderia pseudomallei type III secretion system (T3SS), has been overexpressed as a GST fusion, cleaved from the GST tag and purified. Crystals were grown of native and selenomethionine-labelled BipD. The crystals grow in two different polymorphs from the same condition. The first polymorph belongs to space group C222, with unit-cell parameters a = 103.98, b = 122.79, c = 49.17 A, a calculated Matthews coefficient of 2.4 A(3) Da(-1) (47% solvent content) and one molecule per asymmetric unit. The second polymorph belongs to space group P2(1)2(1)2, with unit-cell parameters a = 136.47, b = 89.84, c = 50.15 A, and a calculated Matthews coefficient of 2.3 A(3) Da(-1) (45% solvent content) for two molecules per asymmetric unit (analysis of the self-rotation function indicates the presence of a weak twofold non-crystallographic symmetry axis in this P2(1)2(1)2 form). The native crystals of both forms give diffraction data to 2.7 A resolution, while the SeMet-labelled P2(1)2(1)2 crystals diffract to 3.3 A resolution. A K(2)PtCl(4) derivative of the P2(1)2(1)2 form was also obtained and data were collected to 2.7 A with radiation of wavelength lambda = 0.933 A. The Pt-derivative anomalous difference Patterson map revealed two self-peaks on the Harker sections. |
| spellingShingle | Roversi, P Johnson, S Field, T Deane, J Galyov, E Lea, S Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title | Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title_full | Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title_fullStr | Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title_full_unstemmed | Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title_short | Expression, purification, crystallization and preliminary crystallographic analysis of BipD, a component of the Burkholderia pseudomallei type III secretion system. |
| title_sort | expression purification crystallization and preliminary crystallographic analysis of bipd a component of the burkholderia pseudomallei type iii secretion system |
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