Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies

The 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...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:PLoS pathogens 2019-10, Vol.15 (10), p.e1008117-e1008117
Hauptverfasser: Eraña, Hasier, Charco, Jorge M, Di Bari, Michele A, Díaz-Domínguez, Carlos M, López-Moreno, Rafael, Vidal, Enric, González-Miranda, Ezequiel, Pérez-Castro, Miguel A, García-Martínez, Sandra, Bravo, Susana, Fernández-Borges, Natalia, Geijo, Mariví, D'Agostino, Claudia, Garrido, Joseba, Bian, Jifeng, König, Anna, Uluca-Yazgi, Boran, Sabate, Raimon, Khaychuk, Vadim, Vanni, Ilaria, Telling, Glenn C, Heise, Henrike, Nonno, Romolo, Requena, Jesús R, Castilla, Joaquín
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page e1008117
container_issue 10
container_start_page e1008117
container_title PLoS pathogens
container_volume 15
creator Eraña, Hasier
Charco, Jorge M
Di Bari, Michele A
Díaz-Domínguez, Carlos M
López-Moreno, Rafael
Vidal, Enric
González-Miranda, Ezequiel
Pérez-Castro, Miguel A
García-Martínez, Sandra
Bravo, Susana
Fernández-Borges, Natalia
Geijo, Mariví
D'Agostino, Claudia
Garrido, Joseba
Bian, Jifeng
König, Anna
Uluca-Yazgi, Boran
Sabate, Raimon
Khaychuk, Vadim
Vanni, Ilaria
Telling, Glenn C
Heise, Henrike
Nonno, Romolo
Requena, Jesús R
Castilla, Joaquín
description The 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.
doi_str_mv 10.1371/journal.ppat.1008117
format Article
fullrecord <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_2314933907</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A604840201</galeid><doaj_id>oai_doaj_org_article_215467d1d4f44efeb809d884605f8666</doaj_id><sourcerecordid>A604840201</sourcerecordid><originalsourceid>FETCH-LOGICAL-c664t-1950874cfcda28df22e14d0d8e14bb90213e7a3ec89245143491cc72529a87ba3</originalsourceid><addsrcrecordid>eNqVU8uO1DAQjBCIXRb-AEEkLnCYwa_EzgVptbxGWoHE42w5TieTkRMH2xnY3-FLcTLZ0Q7iwiF2O11V7S6rk-QpRmtMOX69s6PrlVkPgwprjJDAmN9LznGW0RWnnN2_E58lj7zfIcQwxfnD5CyujGWcnSe_38IejB066ENq61SlPfxMjXINmJvUa2VUaSBt-3TfBmfTwbW2j6sdVKPCFHcQtrZKa-vSsIUpVY16zkS5tq9hOow-daBtV7a9ioVmFT9ztm2zjTlvzTiTfHCRPjplYjhWLfjHyYNaGQ9Plv0i-f7-3berj6vrzx82V5fXK53nLKxwkSHBma51pYioakIAswpVIm5lWSCCKXBFQYuCsAwzygqsNScZKZTgpaIXyfOD7mCsl4u9XhKKWUFpgXhEbA6IyqqdjE10yt1Iq1o5_7CukcqFVhuQBGcs5xWuWM0Y1FAKVFRCsBxltcjzPGq9WaqNZQeVjv7Hnk9ETzN9u5WN3ctcEF5gEQXwQUD7UcvoLjitwkw8HqaPIE4kzXkhJs7LpaizP0bwQXat12CM6iG-UewViYxM-hH64i_ovx1ZUI2KTcfXtvGuehKVlzligiGCJq1XJyht-wC_QqNG7-Xm65f_wH46xbLFAme9d1Af7cNITkNye2c5DYlchiTSnt21_ki6nQr6B6rrEIs</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2314933907</pqid></control><display><type>article</type><title>Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Recercat</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>PubMed Central Open Access</source><source>Public Library of Science (PLoS)</source><creator>Eraña, Hasier ; Charco, Jorge M ; Di Bari, Michele A ; Díaz-Domínguez, Carlos M ; López-Moreno, Rafael ; Vidal, Enric ; González-Miranda, Ezequiel ; Pérez-Castro, Miguel A ; García-Martínez, Sandra ; Bravo, Susana ; Fernández-Borges, Natalia ; Geijo, Mariví ; D'Agostino, Claudia ; Garrido, Joseba ; Bian, Jifeng ; König, Anna ; Uluca-Yazgi, Boran ; Sabate, Raimon ; Khaychuk, Vadim ; Vanni, Ilaria ; Telling, Glenn C ; Heise, Henrike ; Nonno, Romolo ; Requena, Jesús R ; Castilla, Joaquín</creator><creatorcontrib>Eraña, Hasier ; Charco, Jorge M ; Di Bari, Michele A ; Díaz-Domínguez, Carlos M ; López-Moreno, Rafael ; Vidal, Enric ; González-Miranda, Ezequiel ; Pérez-Castro, Miguel A ; García-Martínez, Sandra ; Bravo, Susana ; Fernández-Borges, Natalia ; Geijo, Mariví ; D'Agostino, Claudia ; Garrido, Joseba ; Bian, Jifeng ; König, Anna ; Uluca-Yazgi, Boran ; Sabate, Raimon ; Khaychuk, Vadim ; Vanni, Ilaria ; Telling, Glenn C ; Heise, Henrike ; Nonno, Romolo ; Requena, Jesús R ; Castilla, Joaquín</creatorcontrib><description>The 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.</description><identifier>ISSN: 1553-7374</identifier><identifier>ISSN: 1553-7366</identifier><identifier>EISSN: 1553-7374</identifier><identifier>DOI: 10.1371/journal.ppat.1008117</identifier><identifier>PMID: 31644574</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>3D printing ; Amplification ; Animal diseases ; Animals ; Arvicolinae ; Atoms ; Biology ; Biology and Life Sciences ; Central Nervous System - pathology ; Cofactors ; Dextran ; Dextran sulfate ; Dextran Sulfate - pharmacology ; Dextrans ; Diagnostic imaging ; Disease ; Disease Models, Animal ; Food safety ; Heterogeneity ; High resolution ; High resolution spectroscopy ; In vitro methods and tests ; Magnetic resonance imaging ; Mammals ; Medicine and Health Sciences ; Mice, Transgenic ; Molecular modelling ; Neuroimaging ; NMR ; Novels ; Nuclear magnetic resonance ; Nuclear Magnetic Resonance, Biomolecular - methods ; Particles ; Partícules (Matèria) ; Pathogenic microorganisms ; Pharmacy ; Physical Sciences ; Polysaccharides ; Prion diseases ; Prion Diseases - pathology ; Prion protein ; Prion Proteins - metabolism ; Prions ; Prions (Proteins) ; Prions - metabolism ; Propagation ; Protein folding ; Protein purification ; Protein Structure, Tertiary ; Proteins ; Proteostasis Deficiencies - pathology ; Public health ; Research and Analysis Methods ; Shaking ; Sonication ; Sulfates ; Supervision ; Transmissible spongiform encephalopathy ; Àtoms</subject><ispartof>PLoS pathogens, 2019-10, Vol.15 (10), p.e1008117-e1008117</ispartof><rights>COPYRIGHT 2019 Public Library of Science</rights><rights>2019 Eraña et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>cc-by (c) Eraña, Hasier et al., 2019 info:eu-repo/semantics/openAccess &lt;a href="http://creativecommons.org/licenses/by/3.0/es"&gt;http://creativecommons.org/licenses/by/3.0/es&lt;/a&gt;</rights><rights>2019 Eraña et al 2019 Eraña et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c664t-1950874cfcda28df22e14d0d8e14bb90213e7a3ec89245143491cc72529a87ba3</citedby><cites>FETCH-LOGICAL-c664t-1950874cfcda28df22e14d0d8e14bb90213e7a3ec89245143491cc72529a87ba3</cites><orcidid>0000-0003-3894-2362 ; 0000-0001-8776-4211 ; 0000-0001-7615-3079 ; 0000-0002-0410-1963 ; 0000-0001-7556-1564 ; 0000-0002-4965-3286 ; 0000-0002-3120-1038 ; 0000-0002-2216-1361 ; 0000-0003-2617-4263 ; 0000-0002-9081-3894 ; 0000-0003-2850-3470 ; 0000-0003-3476-1855 ; 0000-0003-0943-6823 ; 0000-0002-6910-9202</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6827918/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6827918/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,26951,27901,27902,53766,53768,79569,79570</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31644574$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Eraña, Hasier</creatorcontrib><creatorcontrib>Charco, Jorge M</creatorcontrib><creatorcontrib>Di Bari, Michele A</creatorcontrib><creatorcontrib>Díaz-Domínguez, Carlos M</creatorcontrib><creatorcontrib>López-Moreno, Rafael</creatorcontrib><creatorcontrib>Vidal, Enric</creatorcontrib><creatorcontrib>González-Miranda, Ezequiel</creatorcontrib><creatorcontrib>Pérez-Castro, Miguel A</creatorcontrib><creatorcontrib>García-Martínez, Sandra</creatorcontrib><creatorcontrib>Bravo, Susana</creatorcontrib><creatorcontrib>Fernández-Borges, Natalia</creatorcontrib><creatorcontrib>Geijo, Mariví</creatorcontrib><creatorcontrib>D'Agostino, Claudia</creatorcontrib><creatorcontrib>Garrido, Joseba</creatorcontrib><creatorcontrib>Bian, Jifeng</creatorcontrib><creatorcontrib>König, Anna</creatorcontrib><creatorcontrib>Uluca-Yazgi, Boran</creatorcontrib><creatorcontrib>Sabate, Raimon</creatorcontrib><creatorcontrib>Khaychuk, Vadim</creatorcontrib><creatorcontrib>Vanni, Ilaria</creatorcontrib><creatorcontrib>Telling, Glenn C</creatorcontrib><creatorcontrib>Heise, Henrike</creatorcontrib><creatorcontrib>Nonno, Romolo</creatorcontrib><creatorcontrib>Requena, Jesús R</creatorcontrib><creatorcontrib>Castilla, Joaquín</creatorcontrib><title>Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies</title><title>PLoS pathogens</title><addtitle>PLoS Pathog</addtitle><description>The 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.</description><subject>3D printing</subject><subject>Amplification</subject><subject>Animal diseases</subject><subject>Animals</subject><subject>Arvicolinae</subject><subject>Atoms</subject><subject>Biology</subject><subject>Biology and Life Sciences</subject><subject>Central Nervous System - pathology</subject><subject>Cofactors</subject><subject>Dextran</subject><subject>Dextran sulfate</subject><subject>Dextran Sulfate - pharmacology</subject><subject>Dextrans</subject><subject>Diagnostic imaging</subject><subject>Disease</subject><subject>Disease Models, Animal</subject><subject>Food safety</subject><subject>Heterogeneity</subject><subject>High resolution</subject><subject>High resolution spectroscopy</subject><subject>In vitro methods and tests</subject><subject>Magnetic resonance imaging</subject><subject>Mammals</subject><subject>Medicine and Health Sciences</subject><subject>Mice, Transgenic</subject><subject>Molecular modelling</subject><subject>Neuroimaging</subject><subject>NMR</subject><subject>Novels</subject><subject>Nuclear magnetic resonance</subject><subject>Nuclear Magnetic Resonance, Biomolecular - methods</subject><subject>Particles</subject><subject>Partícules (Matèria)</subject><subject>Pathogenic microorganisms</subject><subject>Pharmacy</subject><subject>Physical Sciences</subject><subject>Polysaccharides</subject><subject>Prion diseases</subject><subject>Prion Diseases - pathology</subject><subject>Prion protein</subject><subject>Prion Proteins - metabolism</subject><subject>Prions</subject><subject>Prions (Proteins)</subject><subject>Prions - metabolism</subject><subject>Propagation</subject><subject>Protein folding</subject><subject>Protein purification</subject><subject>Protein Structure, Tertiary</subject><subject>Proteins</subject><subject>Proteostasis Deficiencies - pathology</subject><subject>Public health</subject><subject>Research and Analysis Methods</subject><subject>Shaking</subject><subject>Sonication</subject><subject>Sulfates</subject><subject>Supervision</subject><subject>Transmissible spongiform encephalopathy</subject><subject>Àtoms</subject><issn>1553-7374</issn><issn>1553-7366</issn><issn>1553-7374</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>XX2</sourceid><sourceid>DOA</sourceid><recordid>eNqVU8uO1DAQjBCIXRb-AEEkLnCYwa_EzgVptbxGWoHE42w5TieTkRMH2xnY3-FLcTLZ0Q7iwiF2O11V7S6rk-QpRmtMOX69s6PrlVkPgwprjJDAmN9LznGW0RWnnN2_E58lj7zfIcQwxfnD5CyujGWcnSe_38IejB066ENq61SlPfxMjXINmJvUa2VUaSBt-3TfBmfTwbW2j6sdVKPCFHcQtrZKa-vSsIUpVY16zkS5tq9hOow-daBtV7a9ioVmFT9ztm2zjTlvzTiTfHCRPjplYjhWLfjHyYNaGQ9Plv0i-f7-3berj6vrzx82V5fXK53nLKxwkSHBma51pYioakIAswpVIm5lWSCCKXBFQYuCsAwzygqsNScZKZTgpaIXyfOD7mCsl4u9XhKKWUFpgXhEbA6IyqqdjE10yt1Iq1o5_7CukcqFVhuQBGcs5xWuWM0Y1FAKVFRCsBxltcjzPGq9WaqNZQeVjv7Hnk9ETzN9u5WN3ctcEF5gEQXwQUD7UcvoLjitwkw8HqaPIE4kzXkhJs7LpaizP0bwQXat12CM6iG-UewViYxM-hH64i_ovx1ZUI2KTcfXtvGuehKVlzligiGCJq1XJyht-wC_QqNG7-Xm65f_wH46xbLFAme9d1Af7cNITkNye2c5DYlchiTSnt21_ki6nQr6B6rrEIs</recordid><startdate>20191023</startdate><enddate>20191023</enddate><creator>Eraña, Hasier</creator><creator>Charco, Jorge M</creator><creator>Di Bari, Michele A</creator><creator>Díaz-Domínguez, Carlos M</creator><creator>López-Moreno, Rafael</creator><creator>Vidal, Enric</creator><creator>González-Miranda, Ezequiel</creator><creator>Pérez-Castro, Miguel A</creator><creator>García-Martínez, Sandra</creator><creator>Bravo, Susana</creator><creator>Fernández-Borges, Natalia</creator><creator>Geijo, Mariví</creator><creator>D'Agostino, Claudia</creator><creator>Garrido, Joseba</creator><creator>Bian, Jifeng</creator><creator>König, Anna</creator><creator>Uluca-Yazgi, Boran</creator><creator>Sabate, Raimon</creator><creator>Khaychuk, Vadim</creator><creator>Vanni, Ilaria</creator><creator>Telling, Glenn C</creator><creator>Heise, Henrike</creator><creator>Nonno, Romolo</creator><creator>Requena, Jesús R</creator><creator>Castilla, Joaquín</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISN</scope><scope>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PIMPY</scope><scope>PJZUB</scope><scope>PKEHL</scope><scope>PPXIY</scope><scope>PQEST</scope><scope>PQGLB</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>XX2</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-3894-2362</orcidid><orcidid>https://orcid.org/0000-0001-8776-4211</orcidid><orcidid>https://orcid.org/0000-0001-7615-3079</orcidid><orcidid>https://orcid.org/0000-0002-0410-1963</orcidid><orcidid>https://orcid.org/0000-0001-7556-1564</orcidid><orcidid>https://orcid.org/0000-0002-4965-3286</orcidid><orcidid>https://orcid.org/0000-0002-3120-1038</orcidid><orcidid>https://orcid.org/0000-0002-2216-1361</orcidid><orcidid>https://orcid.org/0000-0003-2617-4263</orcidid><orcidid>https://orcid.org/0000-0002-9081-3894</orcidid><orcidid>https://orcid.org/0000-0003-2850-3470</orcidid><orcidid>https://orcid.org/0000-0003-3476-1855</orcidid><orcidid>https://orcid.org/0000-0003-0943-6823</orcidid><orcidid>https://orcid.org/0000-0002-6910-9202</orcidid></search><sort><creationdate>20191023</creationdate><title>Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies</title><author>Eraña, Hasier ; Charco, Jorge M ; Di Bari, Michele A ; Díaz-Domínguez, Carlos M ; López-Moreno, Rafael ; Vidal, Enric ; González-Miranda, Ezequiel ; Pérez-Castro, Miguel A ; García-Martínez, Sandra ; Bravo, Susana ; Fernández-Borges, Natalia ; Geijo, Mariví ; D'Agostino, Claudia ; Garrido, Joseba ; Bian, Jifeng ; König, Anna ; Uluca-Yazgi, Boran ; Sabate, Raimon ; Khaychuk, Vadim ; Vanni, Ilaria ; Telling, Glenn C ; Heise, Henrike ; Nonno, Romolo ; Requena, Jesús R ; Castilla, Joaquín</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c664t-1950874cfcda28df22e14d0d8e14bb90213e7a3ec89245143491cc72529a87ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>3D printing</topic><topic>Amplification</topic><topic>Animal diseases</topic><topic>Animals</topic><topic>Arvicolinae</topic><topic>Atoms</topic><topic>Biology</topic><topic>Biology and Life Sciences</topic><topic>Central Nervous System - pathology</topic><topic>Cofactors</topic><topic>Dextran</topic><topic>Dextran sulfate</topic><topic>Dextran Sulfate - pharmacology</topic><topic>Dextrans</topic><topic>Diagnostic imaging</topic><topic>Disease</topic><topic>Disease Models, Animal</topic><topic>Food safety</topic><topic>Heterogeneity</topic><topic>High resolution</topic><topic>High resolution spectroscopy</topic><topic>In vitro methods and tests</topic><topic>Magnetic resonance imaging</topic><topic>Mammals</topic><topic>Medicine and Health Sciences</topic><topic>Mice, Transgenic</topic><topic>Molecular modelling</topic><topic>Neuroimaging</topic><topic>NMR</topic><topic>Novels</topic><topic>Nuclear magnetic resonance</topic><topic>Nuclear Magnetic Resonance, Biomolecular - methods</topic><topic>Particles</topic><topic>Partícules (Matèria)</topic><topic>Pathogenic microorganisms</topic><topic>Pharmacy</topic><topic>Physical Sciences</topic><topic>Polysaccharides</topic><topic>Prion diseases</topic><topic>Prion Diseases - pathology</topic><topic>Prion protein</topic><topic>Prion Proteins - metabolism</topic><topic>Prions</topic><topic>Prions (Proteins)</topic><topic>Prions - metabolism</topic><topic>Propagation</topic><topic>Protein folding</topic><topic>Protein purification</topic><topic>Protein Structure, Tertiary</topic><topic>Proteins</topic><topic>Proteostasis Deficiencies - pathology</topic><topic>Public health</topic><topic>Research and Analysis Methods</topic><topic>Shaking</topic><topic>Sonication</topic><topic>Sulfates</topic><topic>Supervision</topic><topic>Transmissible spongiform encephalopathy</topic><topic>Àtoms</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Eraña, Hasier</creatorcontrib><creatorcontrib>Charco, Jorge M</creatorcontrib><creatorcontrib>Di Bari, Michele A</creatorcontrib><creatorcontrib>Díaz-Domínguez, Carlos M</creatorcontrib><creatorcontrib>López-Moreno, Rafael</creatorcontrib><creatorcontrib>Vidal, Enric</creatorcontrib><creatorcontrib>González-Miranda, Ezequiel</creatorcontrib><creatorcontrib>Pérez-Castro, Miguel A</creatorcontrib><creatorcontrib>García-Martínez, Sandra</creatorcontrib><creatorcontrib>Bravo, Susana</creatorcontrib><creatorcontrib>Fernández-Borges, Natalia</creatorcontrib><creatorcontrib>Geijo, Mariví</creatorcontrib><creatorcontrib>D'Agostino, Claudia</creatorcontrib><creatorcontrib>Garrido, Joseba</creatorcontrib><creatorcontrib>Bian, Jifeng</creatorcontrib><creatorcontrib>König, Anna</creatorcontrib><creatorcontrib>Uluca-Yazgi, Boran</creatorcontrib><creatorcontrib>Sabate, Raimon</creatorcontrib><creatorcontrib>Khaychuk, Vadim</creatorcontrib><creatorcontrib>Vanni, Ilaria</creatorcontrib><creatorcontrib>Telling, Glenn C</creatorcontrib><creatorcontrib>Heise, Henrike</creatorcontrib><creatorcontrib>Nonno, Romolo</creatorcontrib><creatorcontrib>Requena, Jesús R</creatorcontrib><creatorcontrib>Castilla, Joaquín</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Canada</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Virology and AIDS Abstracts</collection><collection>Health &amp; Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>ProQuest Central (New)</collection><collection>ProQuest One Academic (New)</collection><collection>Publicly Available Content Database</collection><collection>ProQuest Health &amp; Medical Research Collection</collection><collection>ProQuest One Academic Middle East (New)</collection><collection>ProQuest One Health &amp; Nursing</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Applied &amp; Life Sciences</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>Recercat</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PLoS pathogens</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Eraña, Hasier</au><au>Charco, Jorge M</au><au>Di Bari, Michele A</au><au>Díaz-Domínguez, Carlos M</au><au>López-Moreno, Rafael</au><au>Vidal, Enric</au><au>González-Miranda, Ezequiel</au><au>Pérez-Castro, Miguel A</au><au>García-Martínez, Sandra</au><au>Bravo, Susana</au><au>Fernández-Borges, Natalia</au><au>Geijo, Mariví</au><au>D'Agostino, Claudia</au><au>Garrido, Joseba</au><au>Bian, Jifeng</au><au>König, Anna</au><au>Uluca-Yazgi, Boran</au><au>Sabate, Raimon</au><au>Khaychuk, Vadim</au><au>Vanni, Ilaria</au><au>Telling, Glenn C</au><au>Heise, Henrike</au><au>Nonno, Romolo</au><au>Requena, Jesús R</au><au>Castilla, Joaquín</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies</atitle><jtitle>PLoS pathogens</jtitle><addtitle>PLoS Pathog</addtitle><date>2019-10-23</date><risdate>2019</risdate><volume>15</volume><issue>10</issue><spage>e1008117</spage><epage>e1008117</epage><pages>e1008117-e1008117</pages><issn>1553-7374</issn><issn>1553-7366</issn><eissn>1553-7374</eissn><abstract>The 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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>31644574</pmid><doi>10.1371/journal.ppat.1008117</doi><tpages>33</tpages><orcidid>https://orcid.org/0000-0003-3894-2362</orcidid><orcidid>https://orcid.org/0000-0001-8776-4211</orcidid><orcidid>https://orcid.org/0000-0001-7615-3079</orcidid><orcidid>https://orcid.org/0000-0002-0410-1963</orcidid><orcidid>https://orcid.org/0000-0001-7556-1564</orcidid><orcidid>https://orcid.org/0000-0002-4965-3286</orcidid><orcidid>https://orcid.org/0000-0002-3120-1038</orcidid><orcidid>https://orcid.org/0000-0002-2216-1361</orcidid><orcidid>https://orcid.org/0000-0003-2617-4263</orcidid><orcidid>https://orcid.org/0000-0002-9081-3894</orcidid><orcidid>https://orcid.org/0000-0003-2850-3470</orcidid><orcidid>https://orcid.org/0000-0003-3476-1855</orcidid><orcidid>https://orcid.org/0000-0003-0943-6823</orcidid><orcidid>https://orcid.org/0000-0002-6910-9202</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 1553-7374
ispartof PLoS pathogens, 2019-10, Vol.15 (10), p.e1008117-e1008117
issn 1553-7374
1553-7366
1553-7374
language eng
recordid cdi_plos_journals_2314933907
source MEDLINE; DOAJ Directory of Open Access Journals; Recercat; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; PubMed Central Open Access; Public Library of Science (PLoS)
subjects 3D printing
Amplification
Animal diseases
Animals
Arvicolinae
Atoms
Biology
Biology and Life Sciences
Central Nervous System - pathology
Cofactors
Dextran
Dextran sulfate
Dextran Sulfate - pharmacology
Dextrans
Diagnostic imaging
Disease
Disease Models, Animal
Food safety
Heterogeneity
High resolution
High resolution spectroscopy
In vitro methods and tests
Magnetic resonance imaging
Mammals
Medicine and Health Sciences
Mice, Transgenic
Molecular modelling
Neuroimaging
NMR
Novels
Nuclear magnetic resonance
Nuclear Magnetic Resonance, Biomolecular - methods
Particles
Partícules (Matèria)
Pathogenic microorganisms
Pharmacy
Physical Sciences
Polysaccharides
Prion diseases
Prion Diseases - pathology
Prion protein
Prion Proteins - metabolism
Prions
Prions (Proteins)
Prions - metabolism
Propagation
Protein folding
Protein purification
Protein Structure, Tertiary
Proteins
Proteostasis Deficiencies - pathology
Public health
Research and Analysis Methods
Shaking
Sonication
Sulfates
Supervision
Transmissible spongiform encephalopathy
Àtoms
title Development of a new largely scalable in vitro prion propagation method for the production of infectious recombinant prions for high resolution structural studies
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-18T00%3A03%3A14IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20a%20new%20largely%20scalable%20in%20vitro%20prion%20propagation%20method%20for%20the%20production%20of%20infectious%20recombinant%20prions%20for%20high%20resolution%20structural%20studies&rft.jtitle=PLoS%20pathogens&rft.au=Era%C3%B1a,%20Hasier&rft.date=2019-10-23&rft.volume=15&rft.issue=10&rft.spage=e1008117&rft.epage=e1008117&rft.pages=e1008117-e1008117&rft.issn=1553-7374&rft.eissn=1553-7374&rft_id=info:doi/10.1371/journal.ppat.1008117&rft_dat=%3Cgale_plos_%3EA604840201%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2314933907&rft_id=info:pmid/31644574&rft_galeid=A604840201&rft_doaj_id=oai_doaj_org_article_215467d1d4f44efeb809d884605f8666&rfr_iscdi=true