Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/103319 https://doi.org/10.3390/ijms23094502 |
Resumo: | Phenylketonuria (PKU) is a rare metabolic disease caused by variations in a human gene, PAH, encoding phenylalanine hydroxylase (PAH), and the enzyme converting the essential amino acid phenylalanine into tyrosine. Many PKU-causing variations compromise the conformational stability of the encoded enzyme, decreasing or abolishing its catalytic activity, and leading to an elevated concentration of phenylalanine in the blood, which is neurotoxic. Several therapeutic approaches have been developed to treat the more severe manifestations of the disorder, but they are either not entirely effective or difficult to adhere to throughout life. In a search for novel pharmacological chaperones to treat PKU, a lead compound was discovered (compound IV) that exhibited promising in vitro and in vivo chaperoning activity on PAH. The structure of the PAH-IV complex has been reported. Here, using alchemical free energy calculations (AFEC) on the structure of the PAH-IV complex, we design a new generation of compound IV-analogues with a higher affinity for the enzyme. Seventeen novel analogues were synthesized, and thermal shift and isothermal titration calorimetry (ITC) assays were performed to experimentally evaluate their stabilizing effect and their affinity for the enzyme. Most of the new derivatives bind to PAH tighter than lead compound IV and induce a greater thermostabilization of the enzyme upon binding. Importantly, the correspondence between the calculated alchemical binding free energies and the experimentally determined ΔΔGb values is excellent, which supports the use of AFEC to design pharmacological chaperones to treat PKU using the X-ray structure of their complexes with the target PAH enzyme. |
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Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylasephenylketonuriapharmacological chaperoneslead optimizationalchemical free energy calculationsbinding energeticsCalorimetryHumansPhenylalanineProtein FoldingPhenylalanine HydroxylasePhenylketonuriasPhenylketonuria (PKU) is a rare metabolic disease caused by variations in a human gene, PAH, encoding phenylalanine hydroxylase (PAH), and the enzyme converting the essential amino acid phenylalanine into tyrosine. Many PKU-causing variations compromise the conformational stability of the encoded enzyme, decreasing or abolishing its catalytic activity, and leading to an elevated concentration of phenylalanine in the blood, which is neurotoxic. Several therapeutic approaches have been developed to treat the more severe manifestations of the disorder, but they are either not entirely effective or difficult to adhere to throughout life. In a search for novel pharmacological chaperones to treat PKU, a lead compound was discovered (compound IV) that exhibited promising in vitro and in vivo chaperoning activity on PAH. The structure of the PAH-IV complex has been reported. Here, using alchemical free energy calculations (AFEC) on the structure of the PAH-IV complex, we design a new generation of compound IV-analogues with a higher affinity for the enzyme. Seventeen novel analogues were synthesized, and thermal shift and isothermal titration calorimetry (ITC) assays were performed to experimentally evaluate their stabilizing effect and their affinity for the enzyme. Most of the new derivatives bind to PAH tighter than lead compound IV and induce a greater thermostabilization of the enzyme upon binding. Importantly, the correspondence between the calculated alchemical binding free energies and the experimentally determined ΔΔGb values is excellent, which supports the use of AFEC to design pharmacological chaperones to treat PKU using the X-ray structure of their complexes with the target PAH enzyme.2022-04-19info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/103319http://hdl.handle.net/10316/103319https://doi.org/10.3390/ijms23094502eng1422-0067Conde-Giménez, MaríaGalano-Frutos, Juan JoséGaliana-Cameo, MaríaMahía, AlejandroVictor, Bruno L.Salillas, SandraVelázquez-Campoy, AdriánBrito, Rui M. M.Gálvez, José AntonioDíaz-de-Villegas, María D.Sancho, Javierinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2022-11-04T21:33:29Zoai:estudogeral.uc.pt:10316/103319Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:20:10.591520Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
title |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
spellingShingle |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase Conde-Giménez, María phenylketonuria pharmacological chaperones lead optimization alchemical free energy calculations binding energetics Calorimetry Humans Phenylalanine Protein Folding Phenylalanine Hydroxylase Phenylketonurias |
title_short |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
title_full |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
title_fullStr |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
title_full_unstemmed |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
title_sort |
Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase |
author |
Conde-Giménez, María |
author_facet |
Conde-Giménez, María Galano-Frutos, Juan José Galiana-Cameo, María Mahía, Alejandro Victor, Bruno L. Salillas, Sandra Velázquez-Campoy, Adrián Brito, Rui M. M. Gálvez, José Antonio Díaz-de-Villegas, María D. Sancho, Javier |
author_role |
author |
author2 |
Galano-Frutos, Juan José Galiana-Cameo, María Mahía, Alejandro Victor, Bruno L. Salillas, Sandra Velázquez-Campoy, Adrián Brito, Rui M. M. Gálvez, José Antonio Díaz-de-Villegas, María D. Sancho, Javier |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Conde-Giménez, María Galano-Frutos, Juan José Galiana-Cameo, María Mahía, Alejandro Victor, Bruno L. Salillas, Sandra Velázquez-Campoy, Adrián Brito, Rui M. M. Gálvez, José Antonio Díaz-de-Villegas, María D. Sancho, Javier |
dc.subject.por.fl_str_mv |
phenylketonuria pharmacological chaperones lead optimization alchemical free energy calculations binding energetics Calorimetry Humans Phenylalanine Protein Folding Phenylalanine Hydroxylase Phenylketonurias |
topic |
phenylketonuria pharmacological chaperones lead optimization alchemical free energy calculations binding energetics Calorimetry Humans Phenylalanine Protein Folding Phenylalanine Hydroxylase Phenylketonurias |
description |
Phenylketonuria (PKU) is a rare metabolic disease caused by variations in a human gene, PAH, encoding phenylalanine hydroxylase (PAH), and the enzyme converting the essential amino acid phenylalanine into tyrosine. Many PKU-causing variations compromise the conformational stability of the encoded enzyme, decreasing or abolishing its catalytic activity, and leading to an elevated concentration of phenylalanine in the blood, which is neurotoxic. Several therapeutic approaches have been developed to treat the more severe manifestations of the disorder, but they are either not entirely effective or difficult to adhere to throughout life. In a search for novel pharmacological chaperones to treat PKU, a lead compound was discovered (compound IV) that exhibited promising in vitro and in vivo chaperoning activity on PAH. The structure of the PAH-IV complex has been reported. Here, using alchemical free energy calculations (AFEC) on the structure of the PAH-IV complex, we design a new generation of compound IV-analogues with a higher affinity for the enzyme. Seventeen novel analogues were synthesized, and thermal shift and isothermal titration calorimetry (ITC) assays were performed to experimentally evaluate their stabilizing effect and their affinity for the enzyme. Most of the new derivatives bind to PAH tighter than lead compound IV and induce a greater thermostabilization of the enzyme upon binding. Importantly, the correspondence between the calculated alchemical binding free energies and the experimentally determined ΔΔGb values is excellent, which supports the use of AFEC to design pharmacological chaperones to treat PKU using the X-ray structure of their complexes with the target PAH enzyme. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-04-19 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/103319 http://hdl.handle.net/10316/103319 https://doi.org/10.3390/ijms23094502 |
url |
http://hdl.handle.net/10316/103319 https://doi.org/10.3390/ijms23094502 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1422-0067 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799134095110832128 |