Journal article
X-ray diffraction analysis and in vitro characterization of the UAM2 protein from Oryza sativa
The role of seemingly non-enzymatic proteins in complexes interconverting UDP-arabinopyranose and UDP-arabinofuranose (UDP-arabinosemutases; UAMs) in the plant cytosol remains unknown. To shed light on their function, crystallographic and functional studies of the seemingly non-enzymatic UAM2 protein from Oryza sativa (OsUAM2) were undertaken.
Here, X-ray diffraction data are reported, as well as analysis of the oligomeric state in the crystal and in solution. OsUAM2 crystallizes readily but forms highly radiation-sensitive crystals with limited diffraction power, requiring careful low-dose vector data acquisition. Using size-exclusion chromatography, it is shown that the protein is monomeric in solution.
Finally, limited proteolysis was employed to demonstrate DTT-enhanced proteolytic digestion, indicating the existence of at least one intramolecular disulfide bridge or, alternatively, a requirement for a structural metal ion.
Language: | English |
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Publisher: | International Union of Crystallography |
Year: | 2017 |
Pages: | 241-245 |
ISSN: | 2053230x |
Types: | Journal article |
DOI: | 10.1107/S2053230X17004587 |
ORCIDs: | Welner, Ditte Hededam and 0000-0002-6702-3560 |
Limited proteolysis Reversibly glycosylated polypeptide UDP-arabinopyranose mutase Vector data collection
Amino Acid Sequence Cloning, Molecular Crystallization Crystallography, X-Ray Dithiothreitol Escherichia coli Gene Expression Genetic Vectors Intramolecular Transferases Oryza Plant Proteins Proteolysis Recombinant Fusion Proteins Subtilisin Uridine Diphosphate Sugars X-Ray Diffraction limited proteolysis reversibly glycosylated polypeptide uridine diphosphate arabinose vector data collection