Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)

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...

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Autores principales: Kaintz, Cornelia, Mayer, Rupert L., Jirsa, Franz, Halbwirth, Heidi, Rompel, Annette
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Science B.V 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4364613/
https://www.ncbi.nlm.nih.gov/pubmed/25697959
http://dx.doi.org/10.1016/j.febslet.2015.02.009
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author Kaintz, Cornelia
Mayer, Rupert L.
Jirsa, Franz
Halbwirth, Heidi
Rompel, Annette
author_facet Kaintz, Cornelia
Mayer, Rupert L.
Jirsa, Franz
Halbwirth, Heidi
Rompel, Annette
author_sort Kaintz, Cornelia
collection PubMed
description 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.
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spelling pubmed-43646132015-04-01 Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1) Kaintz, Cornelia Mayer, Rupert L. Jirsa, Franz Halbwirth, Heidi Rompel, Annette FEBS Lett Article 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. Elsevier Science B.V 2015-03-24 /pmc/articles/PMC4364613/ /pubmed/25697959 http://dx.doi.org/10.1016/j.febslet.2015.02.009 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kaintz, Cornelia
Mayer, Rupert L.
Jirsa, Franz
Halbwirth, Heidi
Rompel, Annette
Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title_full Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title_fullStr Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title_full_unstemmed Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title_short Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)
title_sort site-directed mutagenesis around the cua site of a polyphenol oxidase from coreopsis grandiflora (cgaus1)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4364613/
https://www.ncbi.nlm.nih.gov/pubmed/25697959
http://dx.doi.org/10.1016/j.febslet.2015.02.009
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