Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/162947 |
Resumo: | Degenerative diseases of the Central Nervous System (CNS) are the result of the pro- gressive loss of the ability to respond to nerve impulses, most of which still have no cure. One of the biological structures involved in some pathologies is the dopaminergic (DA) system, with different pathways being mediated by G-protein-coupled receptors (GPCRs), such as the case of dopamine receptors (DRs). Mutations in DRs, namely dopamine D2 receptors (D2R), have been associated with dopaminergic dysregulation, which is consequently associated with var- ious neurological and psychiatric diseases. As a result, modulation of this pathway, using, for example, D2R synthetic antagonists, could result in novel treatments and/or improvement of the quality of life of patients. The work in this thesis aimed at the design of peptides able to interact with the intracel- lular D2R/G-protein interface, which might result in the development of peptide-based com- pounds with therapeutic potential. Some of the peptides contain a cell penetrating sequence (CPP) that will allow translocation of the targeting peptides through the cell membrane. In addition, we aimed to biologically assay peptide-D2R interaction using the TRUPATH assay. After several attempts, we prepared four peptides by Solid Phase Peptide Synthesis (SPPS) on a Wang-PEG resin, namely pep_Gs (FNDCRDIIQRMHLRQYELL), pep_GsTAT (YGRK- KRRQRRR-FNDCRDIIQRMHLRQYELL), pep_Gi12 (FDAVTDVIIKNNLKDCGLF) and pep_Gi12TAT (YGRKKRRQRRR-FDAVTDVIIKNNLKDCGLF). After purification by reversed-phase high perfor- mance liquid chromatography (RP-HPLC), the peptides were characterized by electrospray ion- ization mass spectrometry (ESI-MS). The biological activity of the peptides will be evaluated by the TRUPATH assay after setting up and validation of the assay test method in the laboratory. |
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Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2GPCRsD2RG-proteinpeptidesTRUPATHDomínio/Área Científica::Engenharia e Tecnologia::Engenharia QuímicaDegenerative diseases of the Central Nervous System (CNS) are the result of the pro- gressive loss of the ability to respond to nerve impulses, most of which still have no cure. One of the biological structures involved in some pathologies is the dopaminergic (DA) system, with different pathways being mediated by G-protein-coupled receptors (GPCRs), such as the case of dopamine receptors (DRs). Mutations in DRs, namely dopamine D2 receptors (D2R), have been associated with dopaminergic dysregulation, which is consequently associated with var- ious neurological and psychiatric diseases. As a result, modulation of this pathway, using, for example, D2R synthetic antagonists, could result in novel treatments and/or improvement of the quality of life of patients. The work in this thesis aimed at the design of peptides able to interact with the intracel- lular D2R/G-protein interface, which might result in the development of peptide-based com- pounds with therapeutic potential. Some of the peptides contain a cell penetrating sequence (CPP) that will allow translocation of the targeting peptides through the cell membrane. In addition, we aimed to biologically assay peptide-D2R interaction using the TRUPATH assay. After several attempts, we prepared four peptides by Solid Phase Peptide Synthesis (SPPS) on a Wang-PEG resin, namely pep_Gs (FNDCRDIIQRMHLRQYELL), pep_GsTAT (YGRK- KRRQRRR-FNDCRDIIQRMHLRQYELL), pep_Gi12 (FDAVTDVIIKNNLKDCGLF) and pep_Gi12TAT (YGRKKRRQRRR-FDAVTDVIIKNNLKDCGLF). After purification by reversed-phase high perfor- mance liquid chromatography (RP-HPLC), the peptides were characterized by electrospray ion- ization mass spectrometry (ESI-MS). The biological activity of the peptides will be evaluated by the TRUPATH assay after setting up and validation of the assay test method in the laboratory.As doenças degenerativas do Sistema Nervoso Central (SNC) provêm da perda progres- siva da capacidade de resposta aos impulsos nervosos, sendo a maioria ainda incurável. Uma das estruturas biológicas envolvida nalgumas patologias é o sistema dopaminérgico (DA), constituído por diferentes vias, mediadas por recetores acoplados à proteína G (GPCRs), como os recetores de dopamina (DRs). Mutações nos DRs, sobretudo nos recetores de dopamina D2 (D2R), têm sido associadas à desregulação dopaminérgica, que está relacionada com várias doenças neurológicas e psiquiátricas. Assim, a modulação desta via, usando, p. ex., antagonis- tas sintéticos de D2R, poderia resultar em novos tratamentos ou na melhoria da qualidade de vida dos pacientes. O trabalho desenvolvido nesta tese teve como objetivo a formação de péptidos capazes de interagir com a interface intracelular D2R/Proteína G, podendo resultar no desenvolvimento de compostos com base peptídica com potencial terapêutico. Alguns dos péptidos contêm uma sequência de penetração celular (CPP) que permitirá a passagem dos péptidos alvo atra- vés da membrana celular. Adicionalmente, pretendemos avaliar biologicamente a interação péptido-D2R utilizando o ensaio TRUPATH. Após várias tentativas, obtiveram-se quatro péptidos por síntese de peptídeos em fase sólida (SPPS) na resina Wang-PEG, nomeadamente pep_Gs (FNDCRDIIQRMHLRQYELL), pep_GsTAT (YGRKKRRQRRR-FNDCRDIIQRMHLRQYELL), pep_Gi12 (FDAVTDVIIKNNLKDCGLF) e pep_Gi12TAT (YGRKKRRQRR-FDAVTDVIIKNNLKDCGLF). Após purificação por cromatografia lí- quida de alta eficiência de fase reversa (RP-HPLC), os péptidos foram caracterizados por espe- trometria de massa com ionização por electrospray (ESI-MS). A atividade biológica dos pépti- dos irá ser avaliada pelo ensaio TRUPATH após a sua criação e validação no laboratório.Melo, RitaCorreia, JoãoRUNVicente, Inês Teodósio2024-01-31T12:40:31Z2023-112023-11-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/162947enginfo: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:RCAAP2024-03-11T05:46:00Zoai:run.unl.pt:10362/162947Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:59:10.329473Repositório 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 |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
title |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
spellingShingle |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 Vicente, Inês Teodósio GPCRs D2R G-protein peptides TRUPATH Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
title_short |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
title_full |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
title_fullStr |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
title_full_unstemmed |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
title_sort |
Novel peptides aimed at interacting with the intracellular domain of dopamine receptor type 2 |
author |
Vicente, Inês Teodósio |
author_facet |
Vicente, Inês Teodósio |
author_role |
author |
dc.contributor.none.fl_str_mv |
Melo, Rita Correia, João RUN |
dc.contributor.author.fl_str_mv |
Vicente, Inês Teodósio |
dc.subject.por.fl_str_mv |
GPCRs D2R G-protein peptides TRUPATH Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
topic |
GPCRs D2R G-protein peptides TRUPATH Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
description |
Degenerative diseases of the Central Nervous System (CNS) are the result of the pro- gressive loss of the ability to respond to nerve impulses, most of which still have no cure. One of the biological structures involved in some pathologies is the dopaminergic (DA) system, with different pathways being mediated by G-protein-coupled receptors (GPCRs), such as the case of dopamine receptors (DRs). Mutations in DRs, namely dopamine D2 receptors (D2R), have been associated with dopaminergic dysregulation, which is consequently associated with var- ious neurological and psychiatric diseases. As a result, modulation of this pathway, using, for example, D2R synthetic antagonists, could result in novel treatments and/or improvement of the quality of life of patients. The work in this thesis aimed at the design of peptides able to interact with the intracel- lular D2R/G-protein interface, which might result in the development of peptide-based com- pounds with therapeutic potential. Some of the peptides contain a cell penetrating sequence (CPP) that will allow translocation of the targeting peptides through the cell membrane. In addition, we aimed to biologically assay peptide-D2R interaction using the TRUPATH assay. After several attempts, we prepared four peptides by Solid Phase Peptide Synthesis (SPPS) on a Wang-PEG resin, namely pep_Gs (FNDCRDIIQRMHLRQYELL), pep_GsTAT (YGRK- KRRQRRR-FNDCRDIIQRMHLRQYELL), pep_Gi12 (FDAVTDVIIKNNLKDCGLF) and pep_Gi12TAT (YGRKKRRQRRR-FDAVTDVIIKNNLKDCGLF). After purification by reversed-phase high perfor- mance liquid chromatography (RP-HPLC), the peptides were characterized by electrospray ion- ization mass spectrometry (ESI-MS). The biological activity of the peptides will be evaluated by the TRUPATH assay after setting up and validation of the assay test method in the laboratory. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-11 2023-11-01T00:00:00Z 2024-01-31T12:40:31Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/162947 |
url |
http://hdl.handle.net/10362/162947 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
application/pdf |
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RCAAP |
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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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