X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction

Abstract Current enzymatic systems for quantifying glycated hemoglobin are based on the FAD-containing enzyme fructosyl peptide oxidase (FPOX). FPOX has substrate specificity for fructosyl-α N-valyl-histidine derived from proteolytic digestion of the N-terminus of the HbA1c β-chain. This study repor...

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Autores principales:	Tomohisa Shimasaki, Hiromi Yoshida, Shigehiro Kamitori, Koji Sode
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/59f5cd720d744c0bb6f05331ae360482
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spelling	oai:doaj.org-article:59f5cd720d744c0bb6f05331ae3604822021-12-02T12:32:41ZX-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction10.1038/s41598-017-02657-52045-2322https://doaj.org/article/59f5cd720d744c0bb6f05331ae3604822017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02657-5https://doaj.org/toc/2045-2322Abstract Current enzymatic systems for quantifying glycated hemoglobin are based on the FAD-containing enzyme fructosyl peptide oxidase (FPOX). FPOX has substrate specificity for fructosyl-α N-valyl-histidine derived from proteolytic digestion of the N-terminus of the HbA1c β-chain. This study reports the X-ray structures of the wild-type and Asn56Ala (N56A) mutant of Phaeosphaeria nodorum fructosyl peptide oxidase (PnFPOX) to elucidate the residues responsible for the oxidative half-reaction. N56A showed decreased oxidase activity compared to the wild -type, while its dye-mediated dehydrogenase activity was higher than that of wild type. In wild-type PnFPOX, Asn56 forms a hydrogen bond with Lys274, thereby preventing it from forming a salt bridge with Asp54. By contrast, Lys274 of PnFPOX N56A moves toward Asp54, and they approach each other to form a salt bridge at a distance of 2.92–3.35 Å. Site-directed mutagenesis studies and protein channel analysis suggest that Asp54 assists in accepting oxygen properly at the position of the bound water molecule in the main oxygen channel. These results reveal that Asn56 in PnFPOX is essential for maintaining an effective oxygen accession path, and support the role of Asp54 as a gate keeper that cooperates with Lys274 to enable oxygen to reach the active site properly.Tomohisa ShimasakiHiromi YoshidaShigehiro KamitoriKoji SodeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomohisa Shimasaki Hiromi Yoshida Shigehiro Kamitori Koji Sode X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
description	Abstract Current enzymatic systems for quantifying glycated hemoglobin are based on the FAD-containing enzyme fructosyl peptide oxidase (FPOX). FPOX has substrate specificity for fructosyl-α N-valyl-histidine derived from proteolytic digestion of the N-terminus of the HbA1c β-chain. This study reports the X-ray structures of the wild-type and Asn56Ala (N56A) mutant of Phaeosphaeria nodorum fructosyl peptide oxidase (PnFPOX) to elucidate the residues responsible for the oxidative half-reaction. N56A showed decreased oxidase activity compared to the wild -type, while its dye-mediated dehydrogenase activity was higher than that of wild type. In wild-type PnFPOX, Asn56 forms a hydrogen bond with Lys274, thereby preventing it from forming a salt bridge with Asp54. By contrast, Lys274 of PnFPOX N56A moves toward Asp54, and they approach each other to form a salt bridge at a distance of 2.92–3.35 Å. Site-directed mutagenesis studies and protein channel analysis suggest that Asp54 assists in accepting oxygen properly at the position of the bound water molecule in the main oxygen channel. These results reveal that Asn56 in PnFPOX is essential for maintaining an effective oxygen accession path, and support the role of Asp54 as a gate keeper that cooperates with Lys274 to enable oxygen to reach the active site properly.
format	article
author	Tomohisa Shimasaki Hiromi Yoshida Shigehiro Kamitori Koji Sode
author_facet	Tomohisa Shimasaki Hiromi Yoshida Shigehiro Kamitori Koji Sode
author_sort	Tomohisa Shimasaki
title	X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
title_short	X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
title_full	X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
title_fullStr	X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
title_full_unstemmed	X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
title_sort	x-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/59f5cd720d744c0bb6f05331ae360482
work_keys_str_mv	AT tomohisashimasaki xraystructuresoffructosylpeptideoxidasesrevealingresiduesresponsibleforgatingoxygenaccessintheoxidativehalfreaction AT hiromiyoshida xraystructuresoffructosylpeptideoxidasesrevealingresiduesresponsibleforgatingoxygenaccessintheoxidativehalfreaction AT shigehirokamitori xraystructuresoffructosylpeptideoxidasesrevealingresiduesresponsibleforgatingoxygenaccessintheoxidativehalfreaction AT kojisode xraystructuresoffructosylpeptideoxidasesrevealingresiduesresponsibleforgatingoxygenaccessintheoxidativehalfreaction
_version_	1718393979501805568

X-ray structures of fructosyl peptide oxidases revealing residues responsible for gating oxygen access in the oxidative half reaction

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