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Styrene‐Maleic Acid (SMA) Nanodisc Technology: A Novel Approach for Isolation and Purification of the Infectious Prion Protein (PrP Sc )
Author(s) -
Esmaili Mansoore,
Wang Xiongyao,
Tancowny Brian,
Wille Holger,
Overduin Michael
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.629.15
Subject(s) - chemistry , scrapie , biochemistry , nanodisc , blot , negative stain , lipid bilayer , maleic acid , glycosylation , biophysics , prion protein , membrane , biology , electron microscope , medicine , physics , disease , organic chemistry , pathology , optics , copolymer , gene , polymer
Understanding the structure of infectious prion proteins (PrP Sc ) and their arrangement within the native lipid bilayer has been a daunting task. The challenge arose largely due to an extensive use of conventional detergents such as Sarkosyl and phosphotungstic acid (PTA) derivatives during the purification of PrP Sc , which in turn can exclude physiological lipids and induces fibrillization of PrP Sc into amyloid. Hence, studies based on conventional PrP Sc preparations result in a distorted representation of the in vivo PrP Sc ‐lipid interactions. We demonstrated, for the first time, a novel and fast method for the isolation and purification of PrP Sc from Syrian hamster brains infected with the Hyper prion strain using a number of styrene‐maleic acid (SMA) copolymers. Western blots and silver stained SDS‐PAGEs of the samples (highly enriched in lipids) confirm the presence of PrP Sc in different glycosylation states. The samples were further analyzed by negative stain electron microscopy highlighting key features of the detergent‐free purified PrP Sc including 1) fairly ordered arrays of lipid‐PrP Sc fibrils, 2) formation of 2D crystals under certain experimental conditions and lipid:protein ratios, 3) the appearance of the PrP Sc fibrils is similar to those of detergent purified samples. The SMA platform promises new avenues towards an in‐depth structural, biochemical, and pathological characterization of PrP Sc and its assortment of strains. Support or Funding Information This work has been generously supported by Alberta Innovate BioSolutionDifferent oligomeric structures of isolated PrPscSilver stained and western blotting of isolated PrPSc using Nanodisc technologyCoherent interaction of PrPSc and lipid membranesA glimpse of PrPSc prions dissociated from lipid membranes