Native nanodiscs formed by styrene maleic acid copolymer derivatives help recover infectious prion multimers bound to brain-derived lipids.

Autor:	Esmaili M; Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada., Tancowny BP; Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada.; Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada., Wang X; Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada.; Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada., Moses A; Lipidomics Core Facility, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada., Cortez LM; Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada.; Division of Neurology, Department of Medicine, Centre for Prions and Protein Folding Diseases, and Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Alberta, Canada., Sim VL; Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada.; Division of Neurology, Department of Medicine, Centre for Prions and Protein Folding Diseases, and Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Alberta, Canada., Wille H; Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada overduin@ualberta.ca wille@ualberta.ca.; Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, Alberta, Canada., Overduin M; Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada overduin@ualberta.ca wille@ualberta.ca.
Jazyk:	angličtina
Zdroj:	The Journal of biological chemistry [J Biol Chem] 2020 Jun 19; Vol. 295 (25), pp. 8460-8469. Date of Electronic Publication: 2020 May 01.
DOI:	10.1074/jbc.RA119.012348
Abstrakt:	Prions are lipidated proteins that interact with endogenous lipids and metal ions. They also assemble into multimers and propagate into the infectious scrapie form known as PrP Sc The high-resolution structure of the infectious PrP Sc state remains unknown, and its analysis largely relies on detergent-based preparations devoid of endogenous ligands. Here we designed polymers that allow isolation of endogenous membrane:protein assemblies in native nanodiscs without exposure to conventional detergents that destabilize protein structures and induce fibrillization. A set of styrene-maleic acid (SMA) polymers including a methylamine derivative facilitated gentle release of the infectious complexes for resolution of multimers, and a thiol-containing version promoted crystallization. Polymer extraction from brain homogenates from Syrian hamsters infected with Hyper prions and WT mice infected with Rocky Mountain Laboratories prions yielded infectious prion nanoparticles including oligomers and microfilaments bound to lipid vesicles. Lipid analysis revealed the brain phospholipids that associate with prion protofilaments, as well as those that are specifically enriched in prion assemblies captured by the methylamine-modified copolymer. A comparison of the infectivity of PrP Sc attached to SMA lipid particles in mice and hamsters indicated that these amphipathic polymers offer a valuable tool for high-yield production of intact, detergent-free prions that retain in vivo activity. This native prion isolation method provides an avenue for producing relevant prion:lipid targets and potentially other proteins that form multimeric assemblies and fibrils on membranes. Competing Interests: Conflict of interest—M. O. and M. E. have filed a patent on related SMA copolymers and methods. (© 2020 Esmaili et al.)
Databáze:	MEDLINE
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