PMID- 31118288 OWN - NLM STAT- MEDLINE DCOM- 20200325 LR - 20231213 IS - 1091-6490 (Electronic) IS - 0027-8424 (Print) IS - 0027-8424 (Linking) VI - 116 IP - 23 DP - 2019 Jun 4 TI - Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin. PG - 11229-11234 LID - 10.1073/pnas.1902639116 [doi] AB - 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(4)) that partly acts as a pharmacological chaperone. Here we present structures of full-length human PAH (hPAH) both unbound and complexed with BH(4) in the precatalytic state. Crystal structures, solved at 3.18-A resolution, show the interactions between the cofactor and PAH, explaining the negative regulation exerted by BH(4) BH(4) forms several H-bonds with the N-terminal autoregulatory tail but is far from the catalytic Fe(II) Upon BH(4) binding a polar and salt-bridge interaction network links the three PAH domains, explaining the stability conferred by BH(4) Importantly, BH(4) 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(4) reveals a highly dynamic conformation for the tetramers. Structural analyses of the hPAH:BH(4) subunits revealed that the substrate-induced movement of Tyr138 into the active site could be coupled to the displacement of BH(4) 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. FAU - Flydal, Marte Innselset AU - Flydal MI AUID- ORCID: 0000-0002-4070-8367 AD - Department of Biomedicine, University of Bergen, 5009 Bergen, Norway. FAU - Alcorlo-Pages, Martin AU - Alcorlo-Pages M AUID- ORCID: 0000-0001-8337-6203 AD - Department of Crystallography and Structural Biology, Instituto de Quimica-Fisica "Rocasolano," Consejo Superior de Investigaciones Cientificas (CSIC), 28006 Madrid, Spain. FAU - Johannessen, Fredrik Gullaksen AU - Johannessen FG AUID- ORCID: 0000-0003-2739-9771 AD - Department of Biomedicine, University of Bergen, 5009 Bergen, Norway. FAU - Martinez-Caballero, Siseth AU - Martinez-Caballero S AUID- ORCID: 0000-0003-4842-8447 AD - Department of Biomedicine, University of Bergen, 5009 Bergen, Norway. FAU - Skjaerven, Lars AU - Skjaerven L AUID- ORCID: 0000-0002-8176-4891 AD - Department of Biomedicine, University of Bergen, 5009 Bergen, Norway. FAU - Fernandez-Leiro, Rafael AU - Fernandez-Leiro R AUID- ORCID: 0000-0002-7941-0357 AD - Structural Biology Programme, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain. FAU - Martinez, Aurora AU - Martinez A AUID- ORCID: 0000-0003-1643-6506 AD - Department of Biomedicine, University of Bergen, 5009 Bergen, Norway; aurora.martinez@uib.no xjuan@iqfr.csic.es. FAU - Hermoso, Juan A AU - Hermoso JA AUID- ORCID: 0000-0002-1862-8950 AD - Department of Crystallography and Structural Biology, Instituto de Quimica-Fisica "Rocasolano," Consejo Superior de Investigaciones Cientificas (CSIC), 28006 Madrid, Spain; aurora.martinez@uib.no xjuan@iqfr.csic.es. LA - eng SI - PDB/6HYC SI - PDB/6HPO GR - MC_PC_17136/MRC_/Medical Research Council/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190522 PL - United States TA - Proc Natl Acad Sci U S A JT - Proceedings of the National Academy of Sciences of the United States of America JID - 7505876 RN - 0 (Biopterins) RN - EC 1.14.16.1 (Phenylalanine Hydroxylase) RN - EGX657432I (sapropterin) SB - IM MH - Biopterins/*analogs & derivatives/chemistry/genetics MH - Humans MH - Models, Molecular MH - Molecular Dynamics Simulation MH - Mutagenesis, Site-Directed/methods MH - Mutation/genetics MH - Phenylalanine Hydroxylase/*chemistry/genetics MH - Phenylketonurias/genetics PMC - PMC6561269 OTO - NOTNLM OT - X-ray crystallography OT - allosteric regulation OT - cryo-EM OT - human phenylalanine hydroxylase OT - phenylketonuria COIS- The authors declare no conflict of interest. EDAT- 2019/05/24 06:00 MHDA- 2020/03/26 06:00 PMCR- 2019/11/22 CRDT- 2019/05/24 06:00 PHST- 2019/05/24 06:00 [pubmed] PHST- 2020/03/26 06:00 [medline] PHST- 2019/05/24 06:00 [entrez] PHST- 2019/11/22 00:00 [pmc-release] AID - 1902639116 [pii] AID - 201902639 [pii] AID - 10.1073/pnas.1902639116 [doi] PST - ppublish SO - Proc Natl Acad Sci U S A. 2019 Jun 4;116(23):11229-11234. doi: 10.1073/pnas.1902639116. Epub 2019 May 22.