Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact

Detalhes bibliográficos
Autor(a) principal: Ellena, Matías Nahuel
Data de Publicação: 2023
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/76/76133/tde-06032024-085323/
Resumo: The single-particle cryo-electron microscopy (cryo-EM) advances since the Resolution Revolution has allowed the study of a wide range of proteins that were impossible to analyze with such precision with previous techniques and resolution. The recent acquisition of a Titan Krios microscope in Brazil is a remarkable achievement that positions the country as a leader in cryo-EM in Latin America. However, training researchers across the region is essential to optimize and maximize this technology. Collaborative efforts between Chilean and Brazilian research groups can promote widespread access to this methodology, fostering regional scientific development using state-of-the-art techniques. Here, we present a synergistic approach to determine the cryo-EM structure of Concholepas concholepas hemocyanin (CCH), an oxygen-transporting mega-dalton oligomer found in many invertebrates. Hemocyanins are employed as natural immunostimulants with significant biomedical and clinical applications. Understanding their precise molecular mechanisms is essential for targeted improvement. Structural diversity among homologous hemocyanins, particularly regarding glycosylation patterns, might account for their varying efficiency in generating suitable adjuvant effects in immunotherapy. In preclinical studies, Chilean researchers have shown that CCH holds promise for biomedical applications due to its superior stability, solubility, and immunological response induction. However, its structure and amino acid sequence remain unexplored. This collaboration enabled the elucidation of the structure of the CCH, a heterodidecamer with 8 MDa, and D5 symmetry, presenting two distinct monomers called CCHA and CCHB distributed equally along forming a protein with 366 Å height and 316 Å diameter, for the external collar, and almost 60 glycosylation sites distributed internally and externally, that have application for biomedical properties.
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spelling Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impactDeterminação estrutural, por cryo-EM, da hemocianina do molusco Concholepas concholepas com impacto biomédico substancialCryo-EMCryo-EMHemocianinasHemocyaninMega-Dalton proteinMega-Dalton proteínaThe single-particle cryo-electron microscopy (cryo-EM) advances since the Resolution Revolution has allowed the study of a wide range of proteins that were impossible to analyze with such precision with previous techniques and resolution. The recent acquisition of a Titan Krios microscope in Brazil is a remarkable achievement that positions the country as a leader in cryo-EM in Latin America. However, training researchers across the region is essential to optimize and maximize this technology. Collaborative efforts between Chilean and Brazilian research groups can promote widespread access to this methodology, fostering regional scientific development using state-of-the-art techniques. Here, we present a synergistic approach to determine the cryo-EM structure of Concholepas concholepas hemocyanin (CCH), an oxygen-transporting mega-dalton oligomer found in many invertebrates. Hemocyanins are employed as natural immunostimulants with significant biomedical and clinical applications. Understanding their precise molecular mechanisms is essential for targeted improvement. Structural diversity among homologous hemocyanins, particularly regarding glycosylation patterns, might account for their varying efficiency in generating suitable adjuvant effects in immunotherapy. In preclinical studies, Chilean researchers have shown that CCH holds promise for biomedical applications due to its superior stability, solubility, and immunological response induction. However, its structure and amino acid sequence remain unexplored. This collaboration enabled the elucidation of the structure of the CCH, a heterodidecamer with 8 MDa, and D5 symmetry, presenting two distinct monomers called CCHA and CCHB distributed equally along forming a protein with 366 Å height and 316 Å diameter, for the external collar, and almost 60 glycosylation sites distributed internally and externally, that have application for biomedical properties.Os avanços da criomicroscopia eletrônica de partícula única (cryo-EM) desde a Revolução da Resolução permitiram o estudo de uma ampla gama de proteínas que eram impossíveis de analisar com tanta precisão e resolução com técnicas anteriores. A recente aquisição de um microscópio Titan Krios no Brasil é uma conquista notável que posiciona o país como líder em Cryo-EM na América Latina. Mas para otimizar e maximizar o uso desta tecnologia é de grande importância formar pesquisadores em toda a região. Os esforços colaborativos entre grupos de pesquisa chilenos e brasileiros podem promover o acesso generalizado a esta metodologia, fomentando o desenvolvimento científico regional utilizando técnicas de ponta. Aqui apresentamos uma abordagem sinérgica para determinar por Cryo-EM a estrutura da hemocianina Concholepas concholepas (CCH), um oligômero transportador de oxigênio da ordem do mega-dalton encontrado em muitos invertebrados. As hemocianinas são empregadas como imunoestimulantes naturais com aplicações biomédicas e clínicas significativas. Compreender seus mecanismos moleculares precisos é essencial para uma melhoria direcionada. A diversidade estrutural entre as hemocianinas homólogas, particularmente no que diz respeito aos padrões de glicosilação, pode ser responsável pela sua eficiência variável na geração de efeitos adjuvantes adequados na imunoterapia. Pesquisadores chilenos demonstraram que o CCH é promissor para aplicações biomédicas devido à sua estabilidade superior, a sua solubilidade e a sua indução de resposta imunológica em estudos pré-clínicos. No entanto, a sua estrutura e sequência de aminoácidos permanecem inexploradas. Esta colaboração possibilitou a elucidação da estrutura do CCH, um heterodidecámero, com 8 MDa, e simetria D5, apresentando dois monômeros distintos denominados CCHA e CCHB distribuídos igualmente ao longo da proteína com uma altura de 366 Å e diâmetro de 316 Å, para o colar externo, e quase 60 sítios de glicosilação distribuídos interna e externamente, que têm aplicação e propriedades biomédicas.Biblioteca Digitais de Teses e Dissertações da USPAmbrosio, Andre Luis BerteliEllena, Matías Nahuel2023-12-07info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/76/76133/tde-06032024-085323/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2024-08-22T21:45:02Zoai:teses.usp.br:tde-06032024-085323Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-22T21:45:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
Determinação estrutural, por cryo-EM, da hemocianina do molusco Concholepas concholepas com impacto biomédico substancial
title Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
spellingShingle Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
Ellena, Matías Nahuel
Cryo-EM
Cryo-EM
Hemocianinas
Hemocyanin
Mega-Dalton protein
Mega-Dalton proteína
title_short Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
title_full Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
title_fullStr Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
title_full_unstemmed Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
title_sort Structural determination, by cryo-EM, of the hemocyanin from the mollusk Concholepas concholepas with substantial biomedical impact
author Ellena, Matías Nahuel
author_facet Ellena, Matías Nahuel
author_role author
dc.contributor.none.fl_str_mv Ambrosio, Andre Luis Berteli
dc.contributor.author.fl_str_mv Ellena, Matías Nahuel
dc.subject.por.fl_str_mv Cryo-EM
Cryo-EM
Hemocianinas
Hemocyanin
Mega-Dalton protein
Mega-Dalton proteína
topic Cryo-EM
Cryo-EM
Hemocianinas
Hemocyanin
Mega-Dalton protein
Mega-Dalton proteína
description The single-particle cryo-electron microscopy (cryo-EM) advances since the Resolution Revolution has allowed the study of a wide range of proteins that were impossible to analyze with such precision with previous techniques and resolution. The recent acquisition of a Titan Krios microscope in Brazil is a remarkable achievement that positions the country as a leader in cryo-EM in Latin America. However, training researchers across the region is essential to optimize and maximize this technology. Collaborative efforts between Chilean and Brazilian research groups can promote widespread access to this methodology, fostering regional scientific development using state-of-the-art techniques. Here, we present a synergistic approach to determine the cryo-EM structure of Concholepas concholepas hemocyanin (CCH), an oxygen-transporting mega-dalton oligomer found in many invertebrates. Hemocyanins are employed as natural immunostimulants with significant biomedical and clinical applications. Understanding their precise molecular mechanisms is essential for targeted improvement. Structural diversity among homologous hemocyanins, particularly regarding glycosylation patterns, might account for their varying efficiency in generating suitable adjuvant effects in immunotherapy. In preclinical studies, Chilean researchers have shown that CCH holds promise for biomedical applications due to its superior stability, solubility, and immunological response induction. However, its structure and amino acid sequence remain unexplored. This collaboration enabled the elucidation of the structure of the CCH, a heterodidecamer with 8 MDa, and D5 symmetry, presenting two distinct monomers called CCHA and CCHB distributed equally along forming a protein with 366 Å height and 316 Å diameter, for the external collar, and almost 60 glycosylation sites distributed internally and externally, that have application for biomedical properties.
publishDate 2023
dc.date.none.fl_str_mv 2023-12-07
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dc.language.iso.fl_str_mv eng
language eng
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dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
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