title: Contrasting Effects of Two Lipid Cofactors of Prion Replication on the Conformation of the Prion Protein.
abstract: Recent studies introduced two experimental protocols for converting full-length recombinant prion protein (rPrP) purified from E.coli into the infectious prion state (PrPSc) with high infectivity titers. Both protocols employed protein misfolding cyclic amplification (PMCA) for generating PrPSc 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 PrPSc. 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 PrPSc might be fundamentally different for POPG and PE.
authors.affiliation: Center for Biomedical Engineering and Technology, Department of Anatomy and Neurobiology, University of Maryland School of Medicine, Baltimore, MD, 21201, United States of America.
authors.forename: Saurabh
authors.initials: S
authors.lastname: Srivastava
authors.affiliation: Center for Biomedical Engineering and Technology, Department of Anatomy and Neurobiology, University of Maryland School of Medicine, Baltimore, MD, 21201, United States of America.
authors.forename: Ilia V
authors.initials: IV
authors.lastname: Baskakov
mesh_terms: D000818:Animals; D051379:Mice; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D011328:Prions / Q000737:chemistry*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012997:Solvents / Q000737:chemistry; D044366:Transition Temperature
chemical_list: D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D011328:Prions; D012997:Solvents; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol