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dc.contributor.authorEraña, Hasier
dc.contributor.authorCharco, Jorge M.
dc.contributor.authorDi Bar, Michele A.
dc.contributor.authorDíaz-Domínguez, Carlos M.
dc.contributor.authorLópez-Moreno, Rafael
dc.contributor.authorVidal, Enric
dc.contributor.authorGonzález-Miranda, Ezequiel
dc.contributor.authorPérez-Castro, Miguel A.
dc.contributor.authorGarcía-Martínez, Sandra
dc.contributor.authorBravo, Susana
dc.contributor.authorFernández-Borges, Natalia
dc.contributor.authorGeijo, Mariví
dc.contributor.authorD’Agostino, Claudia
dc.contributor.authorGarrido, Joseba
dc.contributor.authorBian, Jifeng
dc.contributor.authorKönig, Anna
dc.contributor.authorUluca-Yazgi, Boran
dc.contributor.authorSabate, Raimon
dc.contributor.authorKhaychuk, Vadim
dc.contributor.authorVanni, Ilaria
dc.contributor.authorTelling, Glenn C.
dc.contributor.authorHeise, Henrike
dc.contributor.authorNonno, Romolo
dc.contributor.authorRequena, Jesús R.
dc.contributor.authorCastilla, Joaquín
dc.contributor.otherProducció Animalca
dc.date.accessioned2020-04-20T09:56:21Z
dc.date.available2020-04-20T09:56:21Z
dc.date.issued2019-10-23
dc.identifier.citationEraña, Hasier, Jorge M. Charco, Michele A. Di Bari, Carlos M. Díaz-Domínguez, Rafael López-Moreno, Enric Vidal, and Ezequiel González-Miranda et al. 2019. "Development Of A New Largely Scalable In Vitro Prion Propagation Method For The Production Of Infectious Recombinant Prions For High Resolution Structural Studies". PLOS Pathogens 15 (10): e1008117. Public Library of Science (PLoS). doi:10.1371/journal.ppat.1008117.ca
dc.identifier.issn1553-7366ca
dc.identifier.urihttp://hdl.handle.net/20.500.12327/701
dc.description.abstractThe resolution of the three-dimensional structure of infectious prions at the atomic level is pivotal to understand the pathobiology of Transmissible Spongiform Encephalopathies (TSE), but has been long hindered due to certain particularities of these proteinaceous pathogens. Difficulties related to their purification from brain homogenates of disease-affected animals were resolved almost a decade ago by the development of in vitro recombinant prion propagation systems giving rise to highly infectious recombinant prions. However, lack of knowledge about the molecular mechanisms of the misfolding event and the complexity of systems such as the Protein Misfolding Cyclic Amplification (PMCA), have limited generating the large amounts of homogeneous recombinant prion preparations required for high-resolution techniques such as solid state Nuclear Magnetic Resonance (ssNMR) imaging. Herein, we present a novel recombinant prion propagation system based on PMCA that substitutes sonication with shaking thereby allowing the production of unprecedented amounts of multi-labeled, infectious recombinant prions. The use of specific cofactors, such as dextran sulfate, limit the structural heterogeneity of the in vitro propagated prions and makes possible, for the first time, the generation of infectious and likely homogeneous samples in sufficient quantities for studies with high-resolution structural techniques as demonstrated by the preliminary ssNMR spectrum presented here. Overall, we consider that this new method named Protein Misfolding Shaking Amplification (PMSA), opens new avenues to finally elucidate the three-dimensional structure of infectious prions.ca
dc.format.extent33ca
dc.language.isoengca
dc.publisherPublic Library of Scienceca
dc.relation.ispartofPLoS Pathogensca
dc.rightsAttribution 4.0 Internationalca
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.titleDevelopment of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studiesca
dc.typeinfo:eu-repo/semantics/articleca
dc.description.versioninfo:eu-repo/semantics/publishedVersionca
dc.rights.accessLevelinfo:eu-repo/semantics/openAccess
dc.embargo.termscapca
dc.relation.projectIDMINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/AGL2015-65046-C2-1-R/ES/Estudio de los mecanismos moleculares implicados en la diversidad y estabilidad de las cepas priónicas/ca
dc.relation.projectIDMINECO/Programa Estatal de fomento de la investigación científica y técnica de excelencia/BFU2013-48436-C2-1-P/ES/Decodificando los priones: elucidación de la base estructural de la infectividad de proteínas/ca
dc.relation.projectIDMINECO-FEDER/Programa Estatal de fomento de la investigación científica y técnica de excelencia/BFU2017-86692-P/ES/ELUCIDACION DE LA ESTRUCTURA DE LOS PRIONES PRPSC RECOMBINANTES/ca
dc.relation.projectIDEC/INTERREG-POCTEFA/EU/EFA-148-16/ /REDPRIONca
dc.subject.udc619ca
dc.identifier.doihttps://doi.org/10.1371/journal.ppat.1008117ca
dc.contributor.groupSanitat Animalca


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