%0 Journal Article %1 gemmeckerPhytoeneDesaturaseOryza2015 %A Gemmecker, Sandra %A Schaub, Patrick %A Koschmieder, Julian %A Brausemann, Anton %A Drepper, Friedel %A Rodriguez-Franco, Marta %A Ghisla, Sandro %A Warscheid, Bettina %A Einsle, Oliver %A Beyer, Peter %C United States %D 2015 %J PloS one %K & Binding,Protein Biopolymers/*chemistry,Cell Conformation,to_read Electron Electrophoresis,Oryza/*enzymology,Oxidoreductases/*chemistry/isolation Gel Membrane/enzymology,Crystallography Polyacrylamide Scanning,Native Spectrometry,Microscopy X-Ray,Mass purification/metabolism,Protein %N 7 %P e0131717 %R 10.1371/journal.pone.0131717 %T Phytoene Desaturase from Oryza Sativa: Oligomeric Assembly, Membrane Association and Preliminary 3D-Analysis. %V 10 %X Recombinant phytoene desaturase (PDS-His6) from rice was purified to near-homogeneity and shown to be enzymatically active in a biphasic, liposome-based assay system. The protein contains FAD as the sole protein-bound redox-cofactor. Benzoquinones, not replaceable by molecular oxygen, serve as a final electron acceptor defining PDS as a 15-cis-phytoene (donor):plastoquinone oxidoreductase. The herbicidal PDS-inhibitor norflurazon is capable of arresting the reaction by stabilizing the intermediary FAD(red), while an excess of the quinone acceptor relieves this blockage, indicating competition. The enzyme requires its homo-oligomeric association for activity. The sum of data collected through gel permeation chromatography, non-denaturing polyacrylamide electrophoresis, chemical cross-linking, mass spectrometry and electron microscopy techniques indicate that the high-order oligomers formed in solution are the basis for an active preparation. Of these, a tetramer consisting of dimers represents the active unit. This is corroborated by our preliminary X-ray structural analysis that also revealed similarities of the protein fold with the sequence-inhomologous bacterial phytoene desaturase CRTI and other oxidoreductases of the GR2-family of flavoproteins. This points to an evolutionary relatedness of CRTI and PDS yielding different carotene desaturation sequences based on homologous protein folds.

@article{gemmeckerPhytoeneDesaturaseOryza2015, abstract = {Recombinant phytoene desaturase (PDS-His6) from rice was purified to near-homogeneity and shown to be enzymatically active in a biphasic, liposome-based assay system. The protein contains FAD as the sole protein-bound redox-cofactor. Benzoquinones, not replaceable by molecular oxygen, serve as a final electron acceptor defining PDS as a 15-cis-phytoene (donor):plastoquinone oxidoreductase. The herbicidal PDS-inhibitor norflurazon is capable of arresting the reaction by stabilizing the intermediary FAD(red), while an excess of the quinone acceptor relieves this blockage, indicating competition. The enzyme requires its homo-oligomeric association for activity. The sum of data collected through gel permeation chromatography, non-denaturing polyacrylamide electrophoresis, chemical cross-linking, mass spectrometry and electron microscopy techniques indicate that the high-order oligomers formed in solution are the basis for an active preparation. Of these, a tetramer consisting of dimers represents the active unit. This is corroborated by our preliminary X-ray structural analysis that also revealed similarities of the protein fold with the sequence-inhomologous bacterial phytoene desaturase CRTI and other oxidoreductases of the GR2-family of flavoproteins. This points to an evolutionary relatedness of CRTI and PDS yielding different carotene desaturation sequences based on homologous protein folds.}, added-at = {2024-05-17T13:01:35.000+0200}, address = {United States}, author = {Gemmecker, Sandra and Schaub, Patrick and Koschmieder, Julian and Brausemann, Anton and Drepper, Friedel and {Rodriguez-Franco}, Marta and Ghisla, Sandro and Warscheid, Bettina and Einsle, Oliver and Beyer, Peter}, biburl = {https://www.bibsonomy.org/bibtex/253f9bb4bc52bfdb1880e1f788620cebe/warscheidlab}, doi = {10.1371/journal.pone.0131717}, interhash = {b33a99273fdb57a04adf7b1abca7adf8}, intrahash = {53f9bb4bc52bfdb1880e1f788620cebe}, issn = {1932-6203}, journal = {PloS one}, keywords = {& Binding,Protein Biopolymers/*chemistry,Cell Conformation,to_read Electron Electrophoresis,Oryza/*enzymology,Oxidoreductases/*chemistry/isolation Gel Membrane/enzymology,Crystallography Polyacrylamide Scanning,Native Spectrometry,Microscopy X-Ray,Mass purification/metabolism,Protein}, langid = {english}, number = 7, pages = {e0131717}, pmcid = {PMC4492965}, pmid = {26147209}, timestamp = {2024-05-17T13:01:35.000+0200}, title = {Phytoene {{Desaturase}} from {{Oryza}} Sativa: {{Oligomeric Assembly}}, {{Membrane Association}} and {{Preliminary 3D-Analysis}}.}, volume = 10, year = 2015 }