{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T19:56:04Z","timestamp":1770753364003,"version":"3.50.0"},"reference-count":41,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,8]],"date-time":"2022-03-08T00:00:00Z","timestamp":1646697600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UID\/QUI\/50006\/2020 (Associate Laboratory for Green Chemistry\u2013LAQV), UIDB\/04138\/2020 and UIDP\/04138\/2020 (Research Institute for Medicines; iMed.Ulisboa),   UIDB\/04612\/2020 and UIDP\/04612\/2020 (MOSTMICRO- ITQB)"],"award-info":[{"award-number":["UID\/QUI\/50006\/2020 (Associate Laboratory for Green Chemistry\u2013LAQV), UIDB\/04138\/2020 and UIDP\/04138\/2020 (Research Institute for Medicines; iMed.Ulisboa),   UIDB\/04612\/2020 and UIDP\/04612\/2020 (MOSTMICRO- ITQB)"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nanomaterials"],"abstract":"<jats:p>Phenylketonuria (PKU) is an autosomal recessive disease caused by deficient activity of human phenylalanine hydroxylase (hPAH), which can lead to neurologic impairments in untreated patients. Although some therapies are already available for PKU, these are not without drawbacks. Enzyme-replacement therapy through the delivery of functional hPAH could be a promising strategy. In this work, biophysical methods were used to evaluate the potential of [N1112(OH)][C4F9SO3], a biocompatible fluorinated ionic liquid (FIL), as a delivery system of hPAH. The results herein presented show that [N1112(OH)][C4F9SO3] spontaneously forms micelles in a solution that can encapsulate hPAH. This FIL has no significant effect on the secondary structure of hPAH and is able to increase its enzymatic activity, despite the negative impact on protein thermostability. The influence of [N1112(OH)][C4F9SO3] on the complex oligomerization equilibrium of hPAH was also assessed.<\/jats:p>","DOI":"10.3390\/nano12060893","type":"journal-article","created":{"date-parts":[[2022,3,9]],"date-time":"2022-03-09T01:53:54Z","timestamp":1646790834000},"page":"893","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Impact of Fluorinated Ionic Liquids on Human Phenylalanine Hydroxylase\u2014A Potential Drug Delivery System"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2335-400X","authenticated-orcid":false,"given":"M\u00e1rcia M. S.","family":"Alves","sequence":"first","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"},{"name":"LAQV, REQUIMTE, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa (FCT NOVA), 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2946-9342","authenticated-orcid":false,"given":"Paula","family":"Leandro","sequence":"additional","affiliation":[{"name":"Research Institute for Medicines (iMed.Ulisboa) and Department of Pharmaceutical Sciences and Medicines, Faculdade de Farm\u00e1cia, Universidade de Lisboa, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5664-744X","authenticated-orcid":false,"given":"Haydyn D. T.","family":"Mertens","sequence":"additional","affiliation":[{"name":"European Molecular Biology Lamboratory (EMBL), Hamburg Unit c\/o Deutches Elektronen Synchrotron (DESY), D-22607 Hamburg, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7166-6764","authenticated-orcid":false,"given":"Ana B.","family":"Pereiro","sequence":"additional","affiliation":[{"name":"LAQV, REQUIMTE, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa (FCT NOVA), 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8419-5420","authenticated-orcid":false,"given":"Margarida","family":"Archer","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa (ITQB NOVA), 2780-157 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1881","DOI":"10.1039\/C6MD00358C","article-title":"Ionic liquids as a potential tool for drug delivery systems","volume":"7","author":"Adawiyah","year":"2016","journal-title":"MedChemComm"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Pedro, S.N., Freire, C.S.R., Silvestre, A.J.D., and Freire, M.G. (2020). The role of ionic liquids in the pharmaceutical field: An overview of relevant applications. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21218298"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1858","DOI":"10.1080\/03639045.2017.1349788","article-title":"Choline-versus imidazole-based ionic liquids as functional ingredients in topical delivery systems: Cytotoxicity, solubility, and skin permeation studies","volume":"43","author":"Saraiva","year":"2017","journal-title":"Drug Dev. Ind. Pharm."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"63","DOI":"10.2217\/nnm-2020-0340","article-title":"Recent advances in ionic liquids and nanotechnology for drug delivery","volume":"16","author":"Amaral","year":"2020","journal-title":"Nanomedicine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1007\/s12551-018-0407-6","article-title":"Ionic liquids as biocompatible stabilizers of proteins","volume":"10","author":"Reslan","year":"2018","journal-title":"Biophys. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7132","DOI":"10.1021\/acs.chemrev.6b00562","article-title":"Biological Activity of Ionic Liquids and Their Application in Pharmaceutics and Medicine","volume":"117","author":"Egorova","year":"2017","journal-title":"Chem. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1038\/s42004-020-0302-5","article-title":"Revealing the complexity of ionic liquid\u2013protein interactions through a multi-technique investigation","volume":"3","author":"Clarke","year":"2020","journal-title":"Commun. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1016\/j.molliq.2016.07.076","article-title":"Binding interactions of anesthetic drug with surface active ionic liquid","volume":"222","author":"Pal","year":"2016","journal-title":"J. Mol. Liq."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.molliq.2016.10.142","article-title":"Aggregation behavior of pyrrolidinium-based surface active ionic liquids in H2O-EAN binary solvents","volume":"225","author":"Zhou","year":"2017","journal-title":"J. Mol. Liq."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.jcis.2019.04.095","article-title":"Synthesis and characterization of choline\u2013fatty-acid-based ionic liquids: A new biocompatible surfactant","volume":"551","author":"Ali","year":"2019","journal-title":"J. Colloid Interface Sci."},{"key":"ref_11","first-page":"1983","article-title":"Insights into Protein-Ionic Liquid Interactions Aiming at Macromolecule Delivery Systems","volume":"29","author":"Harada","year":"2018","journal-title":"J. Braz. Chem. Soc."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3296","DOI":"10.1021\/acsomega.7b00578","article-title":"Aggregation and Morphological Aptitude of Drug-Based Ionic Liquids in Aqueous Solution","volume":"2","author":"Singh","year":"2017","journal-title":"ACS Omega"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1016\/j.molliq.2017.11.044","article-title":"Pluronic-SAILs (surface active ionic liquids) mixed micelles as efficient hydrophobic quercetin drug carriers","volume":"249","author":"Singla","year":"2018","journal-title":"J. Mol. Liq."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1016\/j.bbagen.2015.08.022","article-title":"Thermodynamic investigations of protein\u2019s behaviour with ionic liquids in aqueous medium studied by isothermal titration calorimetry","volume":"1860","author":"Bharmoria","year":"2016","journal-title":"Biochim. Biophys. Acta-Gen. Subj."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1021\/la503961h","article-title":"Aggregation behavior and total miscibility of fluorinated ionic liquids in water","volume":"31","author":"Pereiro","year":"2015","journal-title":"Langmuir"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.chemosphere.2018.10.159","article-title":"Human cytotoxicity and octanol\/water partition coefficients of fluorinated ionic liquids","volume":"216","author":"Vieira","year":"2019","journal-title":"Chemosphere"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3733","DOI":"10.1021\/acssuschemeng.8b03653","article-title":"Acute Aquatic Toxicity and Biodegradability of Fluorinated Ionic Liquids","volume":"7","author":"Vieira","year":"2019","journal-title":"ACS Sustain. Chem. Eng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.ijpharm.2017.05.002","article-title":"Fluorinated ionic liquids for protein drug delivery systems: Investigating their impact on the structure and function of lysozyme","volume":"526","author":"Alves","year":"2017","journal-title":"Int. J. Pharm."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"114537","DOI":"10.1016\/j.molliq.2020.114537","article-title":"Insights into the interaction of Bovine Serum Albumin with Surface-Active Ionic Liquids in aqueous solution","volume":"322","author":"Alves","year":"2021","journal-title":"J. Mol. Liq."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"13670","DOI":"10.1021\/ja047713s","article-title":"Thermodynamic characterization of the binding of tetrahydropterins to phenylalanine hydroxylase","volume":"126","author":"Pey","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.1016\/j.bbadis.2011.07.012","article-title":"Natural phenylalanine hydroxylase variants that confer a mild phenotype affect the enzyme\u2019s conformational stability and oligomerization equilibrium","volume":"1812","author":"Cerreto","year":"2011","journal-title":"Biochim. Biophys. Acta-Mol. Basis Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"13615","DOI":"10.1038\/s41598-019-49944-x","article-title":"Structure of full-length wild-type human phenylalanine hydroxylase by small angle X-ray scattering reveals substrate-induced conformational stability","volume":"9","author":"Lopes","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1038\/s41572-021-00267-0","article-title":"Phenylketonuria","volume":"7","author":"Blau","year":"2021","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"787242","DOI":"10.3389\/fnmol.2021.787242","article-title":"Engineering Organoids for in vitro Modeling of Phenylketonuria","volume":"14","author":"Borges","year":"2022","journal-title":"Front. Mol. Neurosci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1002\/iub.1150","article-title":"Critical Review Phenylalanine Hydroxylase: Function, Structure, and Regulation","volume":"65","author":"Flydal","year":"2013","journal-title":"IUBMB Life"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1006\/mgme.2000.2970","article-title":"The V388M Mutation Results in a Kinetic Variant Form of Phenylalanine Hydroxylase","volume":"69","author":"Leandro","year":"2000","journal-title":"Mol. Genet. Metab."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"61022","DOI":"10.1039\/C4RA10306H","article-title":"Phenylalanine iminoboronates as new phenylalanine hydroxylase modulators","volume":"4","author":"Montalbano","year":"2014","journal-title":"RSC Adv."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Lopes, R.R., Tom\u00e9, C.S., Russo, R., Paterna, R., Leandro, J., Candeias, N.R., Gon\u00e7alves, L.M.D., Teixeira, M., Sousa, P.M.F., and Guedes, R.C. (2021). Modulation of human phenylalanine hydroxylase by 3-hydroxyquinolin-2(1h)-one derivatives. Biomolecules, 11.","DOI":"10.3390\/biom11030462"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1002\/bip.20853","article-title":"Protein secondary structure analyses from circular dichroism spectroscopy: Methods and reference databases","volume":"89","author":"Whitmore","year":"2008","journal-title":"Biopolymers"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1093\/protein\/6.4.383","article-title":"Evaluation of secondary structure of proteins from UV circular dichroism spectra using an unsupervised learning neural network","volume":"6","author":"Andrade","year":"1993","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"W668","DOI":"10.1093\/nar\/gkh371","article-title":"DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data","volume":"32","author":"Whitmore","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1107\/S160057671500254X","article-title":"Versatile sample environments and automation for biological solution X-ray scattering experiments at the P12 beamline (PETRA III, DESY)","volume":"48","author":"Blanchet","year":"2015","journal-title":"J. Appl. Crystallogr."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1107\/S1600576717007786","article-title":"ATSAS 2.8: A comprehensive data analysis suite for small-angle scattering from macromolecular solutions","volume":"50","author":"Franke","year":"2017","journal-title":"J. Appl. Crystallogr."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1107\/S0021889803012779","article-title":"PRIMUS: A Windows PC-based system for small-angle scattering data analysis","volume":"36","author":"Konarev","year":"2003","journal-title":"J. Appl. Crystallogr."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"10131","DOI":"10.1074\/jbc.RA119.008294","article-title":"Biophysical characterization of full-length human phenylalanine hydroxylase provides a deeper understanding of its quaternary structure equilibrium","volume":"294","author":"Arturo","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.abb.2012.12.017","article-title":"A new model for allosteric regulation of phenylalanine hydroxylase: Implications for disease and therapeutics","volume":"530","author":"Jaffe","year":"2013","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"40937","DOI":"10.1074\/jbc.M112197200","article-title":"Phosphorylation and mutations of Ser16 in human phenylalanine hydroxylase: Kinetic and structural effects","volume":"277","author":"Miranda","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1093\/chromsci\/bmt076","article-title":"Application of ionic liquids in liquid chromatography and electrodriven separation","volume":"51","author":"Huang","year":"2013","journal-title":"J. Chromatogr. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"11229","DOI":"10.1073\/pnas.1902639116","article-title":"Structure of full-length human phenylalanine hydroxylase in complex with tetrahydrobiopterin","volume":"166","author":"Flydal","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2039","DOI":"10.1093\/hmg\/ddq085","article-title":"Pahenu1 is a mouse model for tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency and promotes analysis of the pharmacological chaperone mechanism in vivo","volume":"19","author":"Gersting","year":"2010","journal-title":"Hum. Mol. Genet."}],"container-title":["Nanomaterials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-4991\/12\/6\/893\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:34:00Z","timestamp":1760135640000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-4991\/12\/6\/893"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,8]]},"references-count":41,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["nano12060893"],"URL":"https:\/\/doi.org\/10.3390\/nano12060893","relation":{},"ISSN":["2079-4991"],"issn-type":[{"value":"2079-4991","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,8]]}}}