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Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein

Recent studies introduced two experimental protocols for converting full-length recombinant prion protein (rPrP) purified from E.coli into the infectious prion state (PrP(Sc)) with high infectivity titers. Both protocols employed protein misfolding cyclic amplification (PMCA) for generating PrP(Sc)...

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Autores principales: Srivastava, Saurabh, Baskakov, Ilia V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4474664/
https://www.ncbi.nlm.nih.gov/pubmed/26090881
http://dx.doi.org/10.1371/journal.pone.0130283
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author Srivastava, Saurabh
Baskakov, Ilia V.
author_facet Srivastava, Saurabh
Baskakov, Ilia V.
author_sort Srivastava, Saurabh
collection PubMed
description Recent studies introduced two experimental protocols for converting full-length recombinant prion protein (rPrP) purified from E.coli into the infectious prion state (PrP(Sc)) with high infectivity titers. Both protocols employed protein misfolding cyclic amplification (PMCA) for generating PrP(Sc) de novo, but used two different lipids, 1-palmitoyl-2-oleolyl-sn-glycero-3-phospho(1’-rac-glycerol) (POPG) or phosphatidylethanolamine (PE), as conversion cofactors. The current study compares the effect of POPG and PE on the physical properties of native, α-helical full-length mouse rPrP under the solvent conditions used for converting rPrP into PrP(Sc). Surprisingly, the effects of POPG and PE on rPrP physical properties, including its conformation, thermodynamic stability, aggregation state and interaction with a lipid, were found to be remarkably different. PE was shown to have minimal, if any, effects on rPrP thermodynamic stability, cooperativity of unfolding, immediate solvent environment or aggregation state. In fact, little evidence indicates that PE interacts with rPrP directly. In contrast, POPG was found to bind to and induce dramatic changes in rPrP structure, including a loss of α-helical conformation and formation of large lipid-protein aggregates that were resistant to partially denaturing conditions. These results suggest that the mechanisms by which lipids assist conversion of rPrP into PrP(Sc) might be fundamentally different for POPG and PE.
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spelling pubmed-44746642015-06-30 Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein Srivastava, Saurabh Baskakov, Ilia V. PLoS One Research Article Recent studies introduced two experimental protocols for converting full-length recombinant prion protein (rPrP) purified from E.coli into the infectious prion state (PrP(Sc)) with high infectivity titers. Both protocols employed protein misfolding cyclic amplification (PMCA) for generating PrP(Sc) de novo, but used two different lipids, 1-palmitoyl-2-oleolyl-sn-glycero-3-phospho(1’-rac-glycerol) (POPG) or phosphatidylethanolamine (PE), as conversion cofactors. The current study compares the effect of POPG and PE on the physical properties of native, α-helical full-length mouse rPrP under the solvent conditions used for converting rPrP into PrP(Sc). Surprisingly, the effects of POPG and PE on rPrP physical properties, including its conformation, thermodynamic stability, aggregation state and interaction with a lipid, were found to be remarkably different. PE was shown to have minimal, if any, effects on rPrP thermodynamic stability, cooperativity of unfolding, immediate solvent environment or aggregation state. In fact, little evidence indicates that PE interacts with rPrP directly. In contrast, POPG was found to bind to and induce dramatic changes in rPrP structure, including a loss of α-helical conformation and formation of large lipid-protein aggregates that were resistant to partially denaturing conditions. These results suggest that the mechanisms by which lipids assist conversion of rPrP into PrP(Sc) might be fundamentally different for POPG and PE. Public Library of Science 2015-06-19 /pmc/articles/PMC4474664/ /pubmed/26090881 http://dx.doi.org/10.1371/journal.pone.0130283 Text en © 2015 Srivastava, Baskakov http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Srivastava, Saurabh
Baskakov, Ilia V.
Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title_full Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title_fullStr Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title_full_unstemmed Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title_short Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein
title_sort contrasting effects of two lipid cofactors of prion replication on the conformation of the prion protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4474664/
https://www.ncbi.nlm.nih.gov/pubmed/26090881
http://dx.doi.org/10.1371/journal.pone.0130283
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