Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
- Author(s)
- Cornelia Kaintz, Rupert L. Mayer, Franz Jirsa, Heidi Halbwirth, Annette Rompel
- Abstract
Abstract Aurone synthase from Coreopsis grandiflora (cgAUS1), catalyzing conversion of butein to sulfuretin in a type-3 copper center, is a rare example of a polyphenol oxidase involved in anabolism. Site-directed mutagenesis around the CuA site of AUS1 was performed, and recombinant enzymes were analyzed by mass spectrometry. Replacement of the coordinating CuA histidines with alanine resulted in the presence of a single copper and loss of diphenolase activity. The thioether bridge-building cysteine and a phenylalanine over the CuA site, exchanged to alanine, have no influence on copper content but appear to play an important role in substrate binding.
- Organisation(s)
- Department of Biophysical Chemistry, Department of Analytical Chemistry, Department of Inorganic Chemistry
- External organisation(s)
- Technische Universität Wien
- Journal
- FEBS Letters
- Volume
- 589
- Pages
- 789-797
- No. of pages
- 9
- ISSN
- 0014-5793
- DOI
- https://doi.org/10.1016/j.febslet.2015.02.009
- Publication date
- 03-2015
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 104004 Chemical biology, 106023 Molecular biology, 103006 Chemical physics
- Keywords
- ASJC Scopus subject areas
- Genetics, Molecular Biology, Biophysics, Structural Biology, Biochemistry, Cell Biology
- Portal url
- https://ucrisportal.univie.ac.at/en/publications/sitedirected-mutagenesis-around-the-cua-site-of-a-polyphenol-oxidase-from-coreopsis-grandiflora-cgaus1(e89f1cea-72ce-4adf-a41c-4df008c345f5).html