Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals

13 pages, 12 figures.-- This article is licensed under CC-BY-NC-ND 4.0

Detalhes bibliográficos
Autores: Ferreira, Joana, Domínguez-Arca, Vicente, Carneiro, Joao, Prieto, Gerardo, Taboada, Pablo, Moreira de Campos, Joao
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2024
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/359852
Acesso em linha:http://hdl.handle.net/10261/359852
Access Level:Acceso aberto
Palavra-chave:Crystallization
Crystals
Hydrodynamics¡
Nucleation
Peptides and proteins
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spelling Classical and Nonclassical Nucleation Mechanisms of Insulin CrystalsFerreira, JoanaDomínguez-Arca, VicenteCarneiro, JoaoPrieto, GerardoTaboada, PabloMoreira de Campos, JoaoCrystallizationCrystalsHydrodynamics¡NucleationPeptides and proteins13 pages, 12 figures.-- This article is licensed under CC-BY-NC-ND 4.0Although the Classical Nucleation Theory (CNT) is the most consensual theory to explain protein nucleation mechanisms, experimental observations during the shear-induced assays suggest that the CNT does not always describe the insulin nucleation process. This is the case at intermediate precipitant (ZnCl2) solution concentrations (2.3 mM) and high-temperature values (20 and 40 °C) as well as at low precipitant solution concentrations (1.6 mM) and low-temperature values (5 °C). In this work, crystallization events following the CNT registered at high precipitant solution concentrations (3.1 and 4.7 mM) are typically described by a Newtonian response. On the other hand, crystallization events following a nonclassical nucleation pathway seem to involve the formation of a metastable intermediate state before crystal formation and are described by a transition from Newtonian to shear-thinning responses. A dominant shear-thinning behavior (shear viscosity values ranging more than 6 orders of magnitude) is found during aggregation/agglomeration events. The rheological analysis is complemented with different characterization techniques (Dynamic Light Scattering, Energy-Dispersive Spectroscopy, Circular Dichroism, and Differential Scanning Calorimetry) to understand the insulin behavior in solution, especially during the occurrence of aggregation/agglomeration events. To the best of our knowledge, the current work is the first study describing nonclassical nucleation mechanisms during shear-induced crystallization experiments, which reveals the potential of the interdisciplinary approach herein described and opens a window for a clear understanding of protein nucleation mechanismsJ.F. acknowledges funding from CEFT under FCT/MCTES (PIDDAC) through a postdoctoral scholarship. This work was financially supported by HealthyWaters (NORTE-01-0145-FEDER-000069), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and LA/P/0045/2020 (ALiCE), UIDB/00532/2020, and UIDP/00532/2020 (CEFT), funded by national funds through FCT/MCTES (PIDDAC)Peer reviewedAmerican Chemical SocietyFundação para a Ciência e a Tecnologia (Portugal)European CommissionFerreira, Joana [0000-0002-7069-2693]Domínguez-Arca, Vicente [0000-0002-3500-5915]Prieto, Gerardo [0000-0002-3010-7197]Taboada, Pablo [0000-0002-2903-7857]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/359852reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésFerreira, Joana; Domínguez-Arca, Vicente; Carneiro, Joao; Prieto, Gerardo; Taboada, Pablo; Moreira de Campos, Joao; 2024; Classical and non-classical nucleation mechanisms of insulin crystals supporting information [Dataset]; American Chemical Society; https://doi.org/10.1021/acsomega.3c10052https://doi.org/10.1021/acsomega.3c10052Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3598522026-05-22T06:33:51Z
dc.title.none.fl_str_mv Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
title Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
spellingShingle Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
Ferreira, Joana
Crystallization
Crystals
Hydrodynamics¡
Nucleation
Peptides and proteins
title_short Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
title_full Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
title_fullStr Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
title_full_unstemmed Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
title_sort Classical and Nonclassical Nucleation Mechanisms of Insulin Crystals
dc.creator.none.fl_str_mv Ferreira, Joana
Domínguez-Arca, Vicente
Carneiro, Joao
Prieto, Gerardo
Taboada, Pablo
Moreira de Campos, Joao
author Ferreira, Joana
author_facet Ferreira, Joana
Domínguez-Arca, Vicente
Carneiro, Joao
Prieto, Gerardo
Taboada, Pablo
Moreira de Campos, Joao
author_role author
author2 Domínguez-Arca, Vicente
Carneiro, Joao
Prieto, Gerardo
Taboada, Pablo
Moreira de Campos, Joao
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Fundação para a Ciência e a Tecnologia (Portugal)
European Commission
Ferreira, Joana [0000-0002-7069-2693]
Domínguez-Arca, Vicente [0000-0002-3500-5915]
Prieto, Gerardo [0000-0002-3010-7197]
Taboada, Pablo [0000-0002-2903-7857]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Crystallization
Crystals
Hydrodynamics¡
Nucleation
Peptides and proteins
topic Crystallization
Crystals
Hydrodynamics¡
Nucleation
Peptides and proteins
description 13 pages, 12 figures.-- This article is licensed under CC-BY-NC-ND 4.0
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/359852
url http://hdl.handle.net/10261/359852
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Ferreira, Joana; Domínguez-Arca, Vicente; Carneiro, Joao; Prieto, Gerardo; Taboada, Pablo; Moreira de Campos, Joao; 2024; Classical and non-classical nucleation mechanisms of insulin crystals supporting information [Dataset]; American Chemical Society; https://doi.org/10.1021/acsomega.3c10052
https://doi.org/10.1021/acsomega.3c10052
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.81155

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