Expression, purification, crystallization and preliminary X-ray diffraction analysis of the apo form of InsP 5 2-K from Arabidopsis thaliana

Bãos-Sanz, Jose Ignacio, Sanz-Aparicio, Julia, Brearley, Charles A. ORCID: https://orcid.org/0000-0001-6179-9109 and González, Beatriz (2012) Expression, purification, crystallization and preliminary X-ray diffraction analysis of the apo form of InsP 5 2-K from Arabidopsis thaliana. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 68 (6). pp. 701-704. ISSN 1744-3091

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Abstract

Inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IP 5 2-K) is a key enzyme that catalyzes the synthesis of phytic acid (IP 6) from inositol 1,3,4,5,6-pentakisphos-phate (IP 5) and ATP. The first structure of IP 5 2-K, that from Arabidopsis thaliana, has been solved previously; it only crystallized in the presence of inositol, either the substrate IP 5 or the product IP 6, 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 IP 5 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 P2 12 12, 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
Uncontrolled Keywords:	inositol kinases,inositol phosphate,ip 2-k,ip,phytic acid,biophysics,structural biology,biochemistry,genetics,condensed matter physics ,/dk/atira/pure/subjectarea/asjc/1300/1304
Faculty \ School:	Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Groups > Plant Sciences
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	22 Oct 2019 09:30
Last Modified:	22 Oct 2022 05:22
URI:	https://ueaeprints.uea.ac.uk/id/eprint/72736
DOI:	10.1107/S1744309112017307

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