Isolation and characterization of a polymerized prion protein.
A polymerized form of recombinant mouse prion protein (mPrP) domain 23-231 [mPrP-(23-231)], designated mPrP-z, was generated at acidic pH (pH 2-5) in the presence of selected concentrations of denaturant (2 M guanidinium chloride or 5 M urea). This isoform of mPrP is stable in acidic solution after removal of denaturant. It can be isolated and purified using reversed-phase HPLC or size-exclusion HPLC. mPrP-z bears structural properties that partially resemble those of scrapie prion. Unlike the native mPrP-(23-231) (mPrP-N), mPrP-z exhibits a high content of beta-sheet structure, as shown by CD spectroscopy, and exists as an oligomer with an approximate molecular mass of 340000 Da, as measured by light scattering. However, similarly to mPrP-N, mPrP-z contains the intact disulphide bond and is sensitive to digestion by proteinase K.
Lu
Bao-Yuan
BY
Research Center for Protein Chemistry, Institute of Molecular Medicine, University of Texas at Houston, Houston, Texas 77030, USA.
Chang
Jui-Yoa
JY
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004220:Disulfides; D019286:Endopeptidase K / Q000378:metabolism; D019791:Guanidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008027:Light; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification*; D011328:Prions / Q000737:chemistry* / Q000302:isolation & purification*; D011487:Protein Conformation; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014466:Ultraviolet Rays; D014508:Urea / Q000494:pharmacology
D004220:Disulfides; D010446:Peptide Fragments; D011328:Prions; D020033:Protein Isoforms; C428437:prion protein (23-231); D014508:Urea; D019286:Endopeptidase K; D019791:Guanidine