Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.

Phenylalanine hydroxylase (PAH) is a key enzyme in the catabolism of phenylalanine, and mutations in this enzyme cause phenylketonuria (PKU), a genetic disorder that leads to brain damage and mental retardation if untreated. Some patients benefit from supplementation with a synthetic formulation of...

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Autores: Flydal, Marte Innselset, Alcorlo-Pagés, Martín, Johannessen, Fredrik Gullaksen, Martínez-Caballero, Siseth, Skjærven, Lars, Fernandez-Leiro, Rafael, Martinez, Aurora, Hermoso, Juan A
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Instituto de Salud Carlos III (ISCIII)
Repositorio:Repisalud
Idioma:inglés
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/23566
Acceso en línea:https://hdl.handle.net/20.500.12105/23566
Access Level:acceso abierto
Palabra clave:X-ray crystallography
allosteric regulation
cryo-EM
human phenylalanine hydroxylase
phenylketonuria
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spelling Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.Flydal, Marte InnselsetAlcorlo-Pagés, MartínJohannessen, Fredrik GullaksenMartínez-Caballero, SisethSkjærven, LarsFernandez-Leiro, RafaelMartinez, AuroraHermoso, Juan AFlydal, Marte InnselsetAlcorlo-Pagés, MartínJohannessen, Fredrik GullaksenMartínez-Caballero, SisethSkjærven, LarsFernandez-Leiro, RafaelMartinez, AuroraHermoso, Juan AX-ray crystallographyallosteric regulationcryo-EMhuman phenylalanine hydroxylasephenylketonuriaPhenylalanine hydroxylase (PAH) is a key enzyme in the catabolism of phenylalanine, and mutations in this enzyme cause phenylketonuria (PKU), a genetic disorder that leads to brain damage and mental retardation if untreated. Some patients benefit from supplementation with a synthetic formulation of the cofactor tetrahydrobiopterin (BH) that partly acts as a pharmacological chaperone. Here we present structures of full-length human PAH (hPAH) both unbound and complexed with BH in the precatalytic state. Crystal structures, solved at 3.18-Å resolution, show the interactions between the cofactor and PAH, explaining the negative regulation exerted by BH BH forms several H-bonds with the N-terminal autoregulatory tail but is far from the catalytic Fe Upon BH binding a polar and salt-bridge interaction network links the three PAH domains, explaining the stability conferred by BH Importantly, BH binding modulates the interaction between subunits, providing information about PAH allostery. Moreover, we also show that the cryo-EM structure of hPAH in absence of BH reveals a highly dynamic conformation for the tetramers. Structural analyses of the hPAH:BH subunits revealed that the substrate-induced movement of Tyr138 into the active site could be coupled to the displacement of BH from the precatalytic toward the active conformation, a molecular mechanism that was supported by site-directed mutagenesis and targeted molecular dynamics simulations. Finally, comparison of the rat and human PAH structures show that hPAH is more dynamic, which is related to amino acid substitutions that enhance the flexibility of hPAH and may increase the susceptibility to PKU-associated mutations.National Academy of SciencesMinisterio de Ciencia e Innovación (España)Norway Regional Health Authority20242024-10-0820192019-06-0420192019-06-04research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.12105/23566reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)InglésengAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 BFU2017-90030-P APUNTANDO A LA RESISTENCIA A ANTIBIOTICOS: BASES ESTRUCTURALES DE LA REGULACION EN PROCESOS ESENCIALES DE REMODELADO DE LA PARED CELULARAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 BFU2017-87316-P COMPLEJOS MACROMOLECULARES EN LOS PROCESOS DE REPLICACION Y REPARACION DEL DNA MITOCONDRIAL: MECANISMOS MOLECULARES Y ESTRUCTURALES MEDIANTE CRIO-EMopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/235662026-06-12T12:43:37Z
dc.title.none.fl_str_mv Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
title Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
spellingShingle Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
Flydal, Marte Innselset
X-ray crystallography
allosteric regulation
cryo-EM
human phenylalanine hydroxylase
phenylketonuria
title_short Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
title_full Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
title_fullStr Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
title_full_unstemmed Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
title_sort Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.
dc.creator.none.fl_str_mv Flydal, Marte Innselset
Alcorlo-Pagés, Martín
Johannessen, Fredrik Gullaksen
Martínez-Caballero, Siseth
Skjærven, Lars
Fernandez-Leiro, Rafael
Martinez, Aurora
Hermoso, Juan A
Flydal, Marte Innselset
Alcorlo-Pagés, Martín
Johannessen, Fredrik Gullaksen
Martínez-Caballero, Siseth
Skjærven, Lars
Fernandez-Leiro, Rafael
Martinez, Aurora
Hermoso, Juan A
author Flydal, Marte Innselset
author_facet Flydal, Marte Innselset
Alcorlo-Pagés, Martín
Johannessen, Fredrik Gullaksen
Martínez-Caballero, Siseth
Skjærven, Lars
Fernandez-Leiro, Rafael
Martinez, Aurora
Hermoso, Juan A
author_role author
author2 Alcorlo-Pagés, Martín
Johannessen, Fredrik Gullaksen
Martínez-Caballero, Siseth
Skjærven, Lars
Fernandez-Leiro, Rafael
Martinez, Aurora
Hermoso, Juan A
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Norway Regional Health Authority
dc.subject.none.fl_str_mv X-ray crystallography
allosteric regulation
cryo-EM
human phenylalanine hydroxylase
phenylketonuria
topic X-ray crystallography
allosteric regulation
cryo-EM
human phenylalanine hydroxylase
phenylketonuria
description Phenylalanine hydroxylase (PAH) is a key enzyme in the catabolism of phenylalanine, and mutations in this enzyme cause phenylketonuria (PKU), a genetic disorder that leads to brain damage and mental retardation if untreated. Some patients benefit from supplementation with a synthetic formulation of the cofactor tetrahydrobiopterin (BH) that partly acts as a pharmacological chaperone. Here we present structures of full-length human PAH (hPAH) both unbound and complexed with BH in the precatalytic state. Crystal structures, solved at 3.18-Å resolution, show the interactions between the cofactor and PAH, explaining the negative regulation exerted by BH BH forms several H-bonds with the N-terminal autoregulatory tail but is far from the catalytic Fe Upon BH binding a polar and salt-bridge interaction network links the three PAH domains, explaining the stability conferred by BH Importantly, BH binding modulates the interaction between subunits, providing information about PAH allostery. Moreover, we also show that the cryo-EM structure of hPAH in absence of BH reveals a highly dynamic conformation for the tetramers. Structural analyses of the hPAH:BH subunits revealed that the substrate-induced movement of Tyr138 into the active site could be coupled to the displacement of BH from the precatalytic toward the active conformation, a molecular mechanism that was supported by site-directed mutagenesis and targeted molecular dynamics simulations. Finally, comparison of the rat and human PAH structures show that hPAH is more dynamic, which is related to amino acid substitutions that enhance the flexibility of hPAH and may increase the susceptibility to PKU-associated mutations.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-06-04
2019
2019-06-04
2024
2024-10-08
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.12105/23566
url https://hdl.handle.net/20.500.12105/23566
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 BFU2017-90030-P APUNTANDO A LA RESISTENCIA A ANTIBIOTICOS: BASES ESTRUCTURALES DE LA REGULACION EN PROCESOS ESENCIALES DE REMODELADO DE LA PARED CELULAR
Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 BFU2017-87316-P COMPLEJOS MACROMOLECULARES EN LOS PROCESOS DE REPLICACION Y REPARACION DEL DNA MITOCONDRIAL: MECANISMOS MOLECULARES Y ESTRUCTURALES MEDIANTE CRIO-EM
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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