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The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis

The UbiD family of reversible decarboxylases act on aromatic, heteroaromatic, and unsaturated aliphatic acids and utilize a prenylated flavin mononucleotide (prFMN) as cofactor, bound adjacent to a conserved Glu–Arg–Glu/Asp ionic network in the enzyme's active site. It is proposed that UbiD act...

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Autores principales: Bailey, Samuel S., Payne, Karl A. P., Fisher, Karl, Marshall, Stephen A., Cliff, Matthew J., Spiess, Reynard, Parker, David A., Rigby, Stephen E. J., Leys, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5818171/
https://www.ncbi.nlm.nih.gov/pubmed/29259125
http://dx.doi.org/10.1074/jbc.RA117.000881
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author Bailey, Samuel S.
Payne, Karl A. P.
Fisher, Karl
Marshall, Stephen A.
Cliff, Matthew J.
Spiess, Reynard
Parker, David A.
Rigby, Stephen E. J.
Leys, David
author_facet Bailey, Samuel S.
Payne, Karl A. P.
Fisher, Karl
Marshall, Stephen A.
Cliff, Matthew J.
Spiess, Reynard
Parker, David A.
Rigby, Stephen E. J.
Leys, David
author_sort Bailey, Samuel S.
collection PubMed
description The UbiD family of reversible decarboxylases act on aromatic, heteroaromatic, and unsaturated aliphatic acids and utilize a prenylated flavin mononucleotide (prFMN) as cofactor, bound adjacent to a conserved Glu–Arg–Glu/Asp ionic network in the enzyme's active site. It is proposed that UbiD activation requires oxidative maturation of the cofactor, for which two distinct isomers, prFMN(ketimine) and prFMN(iminium), have been observed. It also has been suggested that only the prFMN(iminium) form is relevant to catalysis, which requires transient cycloaddition between substrate and cofactor. Using Aspergillus niger Fdc1 as a model system, we reveal that isomerization of prFMN(iminium) to prFMN(ketimine) is a light-dependent process that is largely independent of the Glu(277)–Arg(173)–Glu(282) network and accompanied by irreversible loss of activity. On the other hand, efficient catalysis was highly dependent on an intact Glu–Arg–Glu network, as only Glu → Asp substitutions retain activity. Surprisingly, oxidative maturation to form the prFMN(iminium) species is severely affected only for the R173A variant. In summary, the unusual irreversible isomerization of prFMN is light-dependent and probably proceeds via high-energy intermediates but is independent of the Glu–Arg–Glu network. Our results from mutagenesis, crystallographic, spectroscopic, and kinetic experiments indicate a clear role for the Glu–Arg–Glu network in both catalysis and oxidative maturation.
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spelling pubmed-58181712018-02-21 The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis Bailey, Samuel S. Payne, Karl A. P. Fisher, Karl Marshall, Stephen A. Cliff, Matthew J. Spiess, Reynard Parker, David A. Rigby, Stephen E. J. Leys, David J Biol Chem Enzymology The UbiD family of reversible decarboxylases act on aromatic, heteroaromatic, and unsaturated aliphatic acids and utilize a prenylated flavin mononucleotide (prFMN) as cofactor, bound adjacent to a conserved Glu–Arg–Glu/Asp ionic network in the enzyme's active site. It is proposed that UbiD activation requires oxidative maturation of the cofactor, for which two distinct isomers, prFMN(ketimine) and prFMN(iminium), have been observed. It also has been suggested that only the prFMN(iminium) form is relevant to catalysis, which requires transient cycloaddition between substrate and cofactor. Using Aspergillus niger Fdc1 as a model system, we reveal that isomerization of prFMN(iminium) to prFMN(ketimine) is a light-dependent process that is largely independent of the Glu(277)–Arg(173)–Glu(282) network and accompanied by irreversible loss of activity. On the other hand, efficient catalysis was highly dependent on an intact Glu–Arg–Glu network, as only Glu → Asp substitutions retain activity. Surprisingly, oxidative maturation to form the prFMN(iminium) species is severely affected only for the R173A variant. In summary, the unusual irreversible isomerization of prFMN is light-dependent and probably proceeds via high-energy intermediates but is independent of the Glu–Arg–Glu network. Our results from mutagenesis, crystallographic, spectroscopic, and kinetic experiments indicate a clear role for the Glu–Arg–Glu network in both catalysis and oxidative maturation. American Society for Biochemistry and Molecular Biology 2018-02-16 2017-12-19 /pmc/articles/PMC5818171/ /pubmed/29259125 http://dx.doi.org/10.1074/jbc.RA117.000881 Text en © 2018 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Enzymology
Bailey, Samuel S.
Payne, Karl A. P.
Fisher, Karl
Marshall, Stephen A.
Cliff, Matthew J.
Spiess, Reynard
Parker, David A.
Rigby, Stephen E. J.
Leys, David
The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title_full The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title_fullStr The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title_full_unstemmed The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title_short The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
title_sort role of conserved residues in fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis
topic Enzymology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5818171/
https://www.ncbi.nlm.nih.gov/pubmed/29259125
http://dx.doi.org/10.1074/jbc.RA117.000881
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