Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation

Detalhes bibliográficos
Autor(a) principal: Gonçalves, Ana M.
Data de Publicação: 2022
Outros Autores: Sousa, Ângela, Pedro, Augusto Q., Romão, Maria J., Queiroz, João A., Gallardo, Eugénia, Passarinha, Luís A.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/143977
Resumo: Funding: The authors acknowledge the support from FEDER funds through the POCI—COMPETE 2020—Operational Program Competitiveness and Internationalization in Axis I—Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER-007491) and National Funds (Project UID/Multi/00709/2013) LA/P/0006/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES (PIDDAC). This work was also supported by the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy—i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Researchers also acknowledge funding by FEDER through COMPETE 2020—Programa Operacional Compet-itividade e Internacionalização (POCI), Augusto Q. Pedro research contract CEEC-IND/02599/2020 under the Scientific Stimulus Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
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spelling Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage FormulationDesign of Experimentsenzymatic activityionic liquidsmembrane-bound catechol-O-methyltransferasestabilityCatalysisMolecular BiologySpectroscopyComputer Science ApplicationsPhysical and Theoretical ChemistryOrganic ChemistryInorganic ChemistrySDG 3 - Good Health and Well-beingFunding: The authors acknowledge the support from FEDER funds through the POCI—COMPETE 2020—Operational Program Competitiveness and Internationalization in Axis I—Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER-007491) and National Funds (Project UID/Multi/00709/2013) LA/P/0006/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES (PIDDAC). This work was also supported by the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy—i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Researchers also acknowledge funding by FEDER through COMPETE 2020—Programa Operacional Compet-itividade e Internacionalização (POCI), Augusto Q. Pedro research contract CEEC-IND/02599/2020 under the Scientific Stimulus Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Membrane-bound catechol-O-methyltransferase (MBCOMT), present in the brain and involved in the main pathway of the catechol neurotransmitter deactivation, is linked to several types of human dementia, which are relevant pharmacological targets for new potent and nontoxic inhibitors that have been developed, particularly for Parkinson’s disease treatment. However, the inexistence of an MBCOMT 3D-structure presents a blockage in new drugs’ design and clinical studies due to its instability. The enzyme has a clear tendency to lose its biological activity in a short period of time. To avoid the enzyme sequestering into a non-native state during the downstream processing, a multi-component buffer plays a major role, with the addition of additives such as cysteine, glycerol, and trehalose showing promising results towards minimizing hMBCOMT damage and enhancing its stability. In addition, ionic liquids, due to their virtually unlimited choices for cation/anion paring, are potential protein stabilizers for the process and storage buffers. Screening experiments were designed to evaluate the effect of distinct cation/anion ILs interaction in hMBCOMT enzymatic activity. The ionic liquids: choline glutamate [Ch][Glu], choline dihydrogen phosphate ([Ch][DHP]), choline chloride ([Ch]Cl), 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl), and 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) were supplemented to hMBCOMT lysates in a concentration from 5 to 500 mM. A major potential stabilizing effect was obtained using [Ch][DHP] (10 and 50 mM). From the DoE 146% of hMBCOMT activity recovery was obtained with [Ch][DHP] optimal conditions (7.5 mM) at −80◦ C during 32.4 h. These results are of crucial importance for further drug development once the enzyme can be stabilized for longer periods of time.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaRUNGonçalves, Ana M.Sousa, ÂngelaPedro, Augusto Q.Romão, Maria J.Queiroz, João A.Gallardo, EugéniaPassarinha, Luís A.2022-09-22T22:42:14Z2022-06-302022-06-30T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article14application/pdfhttp://hdl.handle.net/10362/143977eng1661-6596PURE: 46650813https://doi.org/10.3390/ijms23137264info: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:RCAAP2024-03-11T05:23:03Zoai:run.unl.pt:10362/143977Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:51:15.476047Repositório 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 Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
title Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
spellingShingle Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
Gonçalves, Ana M.
Design of Experiments
enzymatic activity
ionic liquids
membrane-bound catechol-O-methyltransferase
stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
title_short Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
title_full Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
title_fullStr Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
title_full_unstemmed Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
title_sort Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation
author Gonçalves, Ana M.
author_facet Gonçalves, Ana M.
Sousa, Ângela
Pedro, Augusto Q.
Romão, Maria J.
Queiroz, João A.
Gallardo, Eugénia
Passarinha, Luís A.
author_role author
author2 Sousa, Ângela
Pedro, Augusto Q.
Romão, Maria J.
Queiroz, João A.
Gallardo, Eugénia
Passarinha, Luís A.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv UCIBIO - Applied Molecular Biosciences Unit
DQ - Departamento de Química
RUN
dc.contributor.author.fl_str_mv Gonçalves, Ana M.
Sousa, Ângela
Pedro, Augusto Q.
Romão, Maria J.
Queiroz, João A.
Gallardo, Eugénia
Passarinha, Luís A.
dc.subject.por.fl_str_mv Design of Experiments
enzymatic activity
ionic liquids
membrane-bound catechol-O-methyltransferase
stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
topic Design of Experiments
enzymatic activity
ionic liquids
membrane-bound catechol-O-methyltransferase
stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
description Funding: The authors acknowledge the support from FEDER funds through the POCI—COMPETE 2020—Operational Program Competitiveness and Internationalization in Axis I—Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER-007491) and National Funds (Project UID/Multi/00709/2013) LA/P/0006/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES (PIDDAC). This work was also supported by the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy—i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Researchers also acknowledge funding by FEDER through COMPETE 2020—Programa Operacional Compet-itividade e Internacionalização (POCI), Augusto Q. Pedro research contract CEEC-IND/02599/2020 under the Scientific Stimulus Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
publishDate 2022
dc.date.none.fl_str_mv 2022-09-22T22:42:14Z
2022-06-30
2022-06-30T00:00:00Z
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/10362/143977
url http://hdl.handle.net/10362/143977
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1661-6596
PURE: 46650813
https://doi.org/10.3390/ijms23137264
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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