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DOI	10.1073/pnas.1902639116
	Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin
	Flydal M.I.; Alcorlo-Pagés M.; Johannessen F.G.; Martínez-Caballero S.; Skjærven L.; Fernandez-Leiro R.; Martinez A.; Hermoso J.A.
发表日期	2019
ISSN	0027-8424
起始页码	11229
结束页码	11234
卷号	166 期号:23
英文摘要	Phenylalanine hydroxylase (PAH) is a key enzyme in the catabo-lism 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 (BH4) that partly acts as a pharmacological chaperone. Here we present structures of full-length human PAH (hPAH) both unbound and complexed with BH4 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 BH4. BH4 forms several H-bonds with the N-terminal autoregulatory tail but is far from the catalytic FeII. Upon BH4 binding a polar and salt-bridge interaction network links the three PAH domains, explaining the stability conferred by BH4. Importantly, BH4 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 BH4 reveals a highly dynamic conformation for the tetramers. Structural analyses of the hPAH:BH4 subunits revealed that the substrate-induced movement of Tyr138 into the active site could be coupled to the displacement of BH4 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. © 2019 National Academy of Sciences. All rights reserved.
英文关键词	Allosteric regulation; Cryo-EM; Human phenylalanine hydroxylase; Phenylketonuria; X-ray crystallography
语种	英语
scopus关键词	ferric ion; phenylalanine 4 monooxygenase; tetrahydrobiopterin; tyrosine; biopterin; phenylalanine 4 monooxygenase; sapropterin; allosterism; amino acid substitution; amino terminal sequence; Article; catalysis; complex formation; conformational transition; controlled study; cryoelectron microscopy; crystal structure; enzyme active site; enzyme analysis; enzyme stability; enzyme structure; enzyme substrate complex; gene mutation; human; hydrogen bond; molecular dynamics; negative feedback; nonhuman; phenylketonuria; priority journal; protein conformation; protein domain; protein protein interaction; rat; site directed mutagenesis; structure analysis; chemistry; genetics; molecular model; mutation; procedures; Biopterin; Humans; Models, Molecular; Molecular Dynamics Simulation; Mutagenesis, Site-Directed; Mutation; Phenylalanine Hydroxylase; Phenylketonurias
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160406
作者单位	Flydal, M.I., Department of Biomedicine, University of Bergen, Bergen, 5009, Norway; Alcorlo-Pagés, M., Department of Crystallography and Structural Biology, Instituto de Química-Física “Rocasolano,”, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, 28006, Spain; Johannessen, F.G., Department of Biomedicine, University of Bergen, Bergen, 5009, Norway; Martínez-Caballero, S., Department of Biomedicine, University of Bergen, Bergen, 5009, Norway; Skjærven, L., Department of Biomedicine, University of Bergen, Bergen, 5009, Norway; Fernandez-Leiro, R., Structural Biology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, 28029, Spain; Martinez, A., Department of Biomedicine, University of Bergen, Bergen, 5009, Norway; Hermoso, J.A., Department of Crystallography and Structural Biology, Instituto de Química-Física “Rocasolano,”, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, 28006, Spain
推荐引用方式 GB/T 7714	Flydal M.I.,Alcorlo-Pagés M.,Johannessen F.G.,et al. Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin[J],2019,166(23).
APA	Flydal M.I..,Alcorlo-Pagés M..,Johannessen F.G..,Martínez-Caballero S..,Skjærven L..,...&Hermoso J.A..(2019).Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin.Proceedings of the National Academy of Sciences of the United States of America,166(23).
MLA	Flydal M.I.,et al."Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin".Proceedings of the National Academy of Sciences of the United States of America 166.23(2019).