Newman, J.A., Das, S.K., Sedelnikova, S.E. et al. (1 more author) (2006) Cloning, purification and preliminary crystallographic analysis of a putative pyridoxal kinase from Bacillus subtilis. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, F62. pp. 1006-1009. ISSN 1744-3091
Abstract
Pyridoxal kinases (PdxK) are able to catalyse the phosphorylation of three vitamin B6 precursors, pyridoxal, pyridoxine and pyridoxamine, to their 5'-phosphates and play an important role in the vitamin B6 salvage pathway. Recently, the thiD gene of Bacillus subtilis was found to encode an enzyme which has the activity expected of a pyridoxal kinase despite its previous assignment as an HMPP kinase owing to higher sequence similarity. As such, this enzyme would appear to represent a new class of `HMPP kinase-like' pyridoxal kinases. B. subtilis thiD has been cloned and the protein has been overexpressed in Escherichia coli, purified and subsequently crystallized in a binary complex with ADP and Mg2+. X-ray diffraction data have been collected from crystals to 2.8 Å resolution at 100 K. The crystals belong to a primitive tetragonal system, point group 422, and analysis of the systematic absences suggest that they belong to one of the enantiomorphic pair of space groups P41212 or P43212. Consideration of the space-group symmetry and unit-cell parameters (a = b = 102.9, c = 252.6 Å, [alpha] = [beta] = [gamma] = 90°) suggest that the crystals contain between three and six molecules in the asymmetric unit. A full structure determination is under way to provide insights into aspects of the enzyme mechanism and substrate specificity.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2006 International Union of Crystallography. Article available Open Access. Article available under the terms of the CC-BY licence. |
Keywords: | thiD; PdxK; HMPP kinase; pyridoxal kinase; ribokinase superfamily |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Infection, Immunity and Cardiovascular Disease The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) > Department of Molecular Biology and Biotechnology (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 06 Jan 2017 11:23 |
Last Modified: | 06 Jan 2017 11:23 |
Published Version: | https://doi.org/10.1107/S1744309106035779 |
Status: | Published |
Publisher: | International Union of Crystallography |
Refereed: | Yes |
Identification Number: | 10.1107/S1744309106035779 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:108133 |