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 protei...

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Veröffentlicht in:Acta crystallographica. Section F, Structural biology communications Structural biology communications, 2017-03, Vol.73 (4)
Hauptverfasser: Welner, Ditte Hededam, Tsai, Alex Yi-Lin, DeGiovanni, Andy M., Scheller, Henrik Vibe, Adams, Paul D.
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container_title Acta crystallographica. Section F, Structural biology communications
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creator Welner, Ditte Hededam
Tsai, Alex Yi-Lin
DeGiovanni, Andy M.
Scheller, Henrik Vibe
Adams, Paul D.
description 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 fromOryza 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.
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subjects BASIC BIOLOGICAL SCIENCES
limited proteolysis
reversibly glycosylated polypeptide
UDP-arabinopyranose mutase
vector data collection
title X-ray diffraction analysis and in vitro characterization of the UAM2 protein from Oryza sativa
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