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Banos-Sanz, J. I., Sanz-Aparicio, J., Brearley, C. A. 
ORCID: https://orcid.org/0000-0001-6179-9109 and Gonzalez, B.
(2012)
Expression, purification, crystallization and preliminary X-ray diffraction analysis of the apo form of Ins5 2-K from Arabidopsis thaliana.
Acta Crystallographica Section F, 68.
pp. 701-704.
Abstract
Inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IP5 2-K) is a key enzyme that catalyzes the synthesis of phytic acid (IP6) from inositol 1,3,4,5,6-pentakisphosphate (IP5) and ATP. The first structure of IP5 2-K, that from Arabidopsis thaliana, has been solved previously; it only crystallized in the presence of inositol, either the substrate IP5 or the product IP6, and failed to crystallize in its free state (without inositol). Based on structural analysis, a point mutation of IP5 2-K (W129A) has been produced in order to overcome this limitation and obtain information about protein conformational changes upon substrate binding. Here, the production and crystallization of W129A IP5 2-K in its free state and with bound nucleotide is described. These crystals differed from the native crystals and belonged to the orthorhombic space group P21212, with unit-cell parameters a = 66.00, b = 68.23, c = 105.80 Å and a = 63.06, b = 71.80, c = 100.23 Å, respectively. The crystals diffracted to resolutions of 2.22 Å (apo) and 2.05 Å (nucleotide bound) using synchrotron radiation and contained one molecule per asymmetric unit. The structures have been determined using the molecular-replacement method and refinement is being undertaken.
| Item Type: | Article |
|---|---|
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Plant Sciences Faculty of Science > Research Groups > Molecular Microbiology |
| Depositing User: | Users 2731 not found. |
| Date Deposited: | 09 Aug 2012 11:57 |
| Last Modified: | 31 Aug 2023 13:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/39316 |
| DOI: | 10.1107/S1744309112017307 |
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