Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity

Protein misfolding is central to numerous neurodegenerative disorders, collectively known as proteinopathies, which include Alzheimer's disease, Parkinson's disease, and prion diseases, among others. In many cases, specific polymorphisms of the proteins associated with these diseases influ...

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Main Authors: Carlos M. Díaz-Domínguez, Hasier Eraña, Francesca Peccati, Enric Vidal, Jorge M. Charco, Cristina Sampedro-Torres-Quevedo, Miguel A. Pérez-Castro, Nuria L. Lorenzo, Samanta Giler, Glenn C. Telling, Mariví Geijo, Jesús R. Requena, Gonzalo Jiménez-Osés, Joaquín Castilla
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Neurobiology of Disease
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Online Access:http://www.sciencedirect.com/science/article/pii/S0969996125002219
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author Carlos M. Díaz-Domínguez
Hasier Eraña
Francesca Peccati
Enric Vidal
Jorge M. Charco
Cristina Sampedro-Torres-Quevedo
Miguel A. Pérez-Castro
Nuria L. Lorenzo
Samanta Giler
Glenn C. Telling
Mariví Geijo
Jesús R. Requena
Gonzalo Jiménez-Osés
Joaquín Castilla
author_facet Carlos M. Díaz-Domínguez
Hasier Eraña
Francesca Peccati
Enric Vidal
Jorge M. Charco
Cristina Sampedro-Torres-Quevedo
Miguel A. Pérez-Castro
Nuria L. Lorenzo
Samanta Giler
Glenn C. Telling
Mariví Geijo
Jesús R. Requena
Gonzalo Jiménez-Osés
Joaquín Castilla
author_sort Carlos M. Díaz-Domínguez
collection DOAJ
description Protein misfolding is central to numerous neurodegenerative disorders, collectively known as proteinopathies, which include Alzheimer's disease, Parkinson's disease, and prion diseases, among others. In many cases, specific polymorphisms of the proteins associated with these diseases influence their misfolding. However, the precise ways in which these polymorphisms affect protein integrity and how they contribute to misfolding propensity remain unclear. In the case of prion diseases, they are caused by prions or PrPSc, the misfolded isoforms of the cellular prion protein (PrPC). Chronic Wasting Disease (CWD) is a prion disease that affects cervids and can exhibit lymphotropic properties, making it the most widespread proteinopathy. For that reason, cervid PrPs and their polymorphisms have been extensively studied. To better understand the role of these polymorphisms, we analyzed 45 cervid PrP variants to assess their effects on flexibility, stability, and spontaneous misfolding propensity.The cervid variants were expressed as recombinant PrP in E. coli and were analyzed for thermal stability using circular dichroism. Additionally, the rec-PrPs served as substrates for Protein Misfolding Shaking Amplification (PMSA), enabling assessment of each variant's spontaneous misfolding propensity. This process led to the formation of bona fide prions, as confirmed by inoculation of one of the resulting conformers into transgenic mice expressing bank vole PrP. In parallel, molecular dynamics simulations were conducted to analyze the structural flexibility of the variants. While differences in protein flexibility were observed, no correlation was detected among flexibility, thermal stability, and the observed variable spontaneous misfolding propensity, suggesting that these properties are independent parameters.
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spelling doaj-art-bd0eb2d41de44854b2b3e7148984575f2025-08-20T03:32:23ZengElsevierNeurobiology of Disease1095-953X2025-09-0121310700510.1016/j.nbd.2025.107005Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensityCarlos M. Díaz-Domínguez0Hasier Eraña1Francesca Peccati2Enric Vidal3Jorge M. Charco4Cristina Sampedro-Torres-Quevedo5Miguel A. Pérez-Castro6Nuria L. Lorenzo7Samanta Giler8Glenn C. Telling9Mariví Geijo10Jesús R. Requena11Gonzalo Jiménez-Osés12Joaquín Castilla13Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Centro de Investigación Biomédica en Red de Enfermedades infecciosas (CIBERINFEC), Carlos III National Health Institute, Madrid, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Centro de Investigación Biomédica en Red de Enfermedades infecciosas (CIBERINFEC), Carlos III National Health Institute, Madrid, Spain.; ATLAS Molecular Pharma S. L., Derio, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Ikerbasque, Basque Foundation for Science, Bilbao, Spain.IRTA. Programa de Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia. Spain.; Unitat mixta d'Investigació IRTA-UAB en Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia. Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Centro de Investigación Biomédica en Red de Enfermedades infecciosas (CIBERINFEC), Carlos III National Health Institute, Madrid, Spain.; ATLAS Molecular Pharma S. L., Derio, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.CIMUS Biomedical Research Institute & Department of Medical Sciences, University of Santiago de Compostela-IDIS, Santiago de Compostela, Spain.IRTA. Programa de Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia. Spain.; Unitat mixta d'Investigació IRTA-UAB en Sanitat Animal. Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Catalonia. Spain.Prion Research Center. Colorado State University, Fort Collins, USA.Animal Health Department, NEIKER-Basque Institute for Agricultural Research and Development, Basque Research and Technology Alliance (BRTA), Derio, Spain.CIMUS Biomedical Research Institute & Department of Medical Sciences, University of Santiago de Compostela-IDIS, Santiago de Compostela, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Ikerbasque, Basque Foundation for Science, Bilbao, Spain.Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.; Centro de Investigación Biomédica en Red de Enfermedades infecciosas (CIBERINFEC), Carlos III National Health Institute, Madrid, Spain.; Ikerbasque, Basque Foundation for Science, Bilbao, Spain.; Corresponding author at: Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.Protein misfolding is central to numerous neurodegenerative disorders, collectively known as proteinopathies, which include Alzheimer's disease, Parkinson's disease, and prion diseases, among others. In many cases, specific polymorphisms of the proteins associated with these diseases influence their misfolding. However, the precise ways in which these polymorphisms affect protein integrity and how they contribute to misfolding propensity remain unclear. In the case of prion diseases, they are caused by prions or PrPSc, the misfolded isoforms of the cellular prion protein (PrPC). Chronic Wasting Disease (CWD) is a prion disease that affects cervids and can exhibit lymphotropic properties, making it the most widespread proteinopathy. For that reason, cervid PrPs and their polymorphisms have been extensively studied. To better understand the role of these polymorphisms, we analyzed 45 cervid PrP variants to assess their effects on flexibility, stability, and spontaneous misfolding propensity.The cervid variants were expressed as recombinant PrP in E. coli and were analyzed for thermal stability using circular dichroism. Additionally, the rec-PrPs served as substrates for Protein Misfolding Shaking Amplification (PMSA), enabling assessment of each variant's spontaneous misfolding propensity. This process led to the formation of bona fide prions, as confirmed by inoculation of one of the resulting conformers into transgenic mice expressing bank vole PrP. In parallel, molecular dynamics simulations were conducted to analyze the structural flexibility of the variants. While differences in protein flexibility were observed, no correlation was detected among flexibility, thermal stability, and the observed variable spontaneous misfolding propensity, suggesting that these properties are independent parameters.http://www.sciencedirect.com/science/article/pii/S0969996125002219CWDMisfoldingPolymorphismsPrionProteinopathyStability
spellingShingle Carlos M. Díaz-Domínguez
Hasier Eraña
Francesca Peccati
Enric Vidal
Jorge M. Charco
Cristina Sampedro-Torres-Quevedo
Miguel A. Pérez-Castro
Nuria L. Lorenzo
Samanta Giler
Glenn C. Telling
Mariví Geijo
Jesús R. Requena
Gonzalo Jiménez-Osés
Joaquín Castilla
Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
Neurobiology of Disease
CWD
Misfolding
Polymorphisms
Prion
Proteinopathy
Stability
title Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
title_full Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
title_fullStr Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
title_full_unstemmed Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
title_short Effect of primary structural variation on cervid prion protein in flexibility, stability, and spontaneous misfolding propensity
title_sort effect of primary structural variation on cervid prion protein in flexibility stability and spontaneous misfolding propensity
topic CWD
Misfolding
Polymorphisms
Prion
Proteinopathy
Stability
url http://www.sciencedirect.com/science/article/pii/S0969996125002219
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