Relative and regional stabilities of the hamster, mouse, rabbit, and bovine prion proteins toward urea unfolding assessed by nuclear magnetic resonance and circular dichroism spectroscopies

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DOIResolve DOI: http://doi.org/10.1021/bi200731e
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TypeArticle
Journal titleBiochemistry
ISSN0006-2960
Volume50
Issue35
Pages75367545; # of pages: 10
SubjectCD spectroscopy; Global stability; High resolution; His-tag; Hydrophobic regions; Mammalian prion proteins; Prion protein; Rank order; Regional stability; Spectroscopic measurements; Dichroism; Mammals; Metabolism; Nuclear magnetic resonance spectroscopy; Proteins; Resonance; Urea; Circular dichroism spectroscopy; polyhistidine tag; prion protein; urea; animal experiment; article; circular dichroism; controlled study; cow; denaturation; hamster; hydrophobicity; mouse; mutation; nonhuman; nuclear magnetic resonance spectroscopy; priority journal; protein purification; proton nuclear magnetic resonance; rabbit; Amino Acid Sequence; Animals; Cattle; Circular Dichroism; Cricetinae; Cricetulus; Mice; Molecular Sequence Data; Nuclear Magnetic Resonance, Biomolecular; Prions; Protein Denaturation; Protein Stability; Protein Unfolding; Rabbits; Species Specificity; Urea; Bovinae; Cricetinae; mammalian prion; Mesocricetus auratus; Oryctolagus cuniculus
AbstractThe residue-specific urea-induced unfolding patterns of recombinant prion proteins from different species (bovine, rabbit, mouse, and Syrian hamster) were monitored using high-resolution 1H nuclear magnetic resonance (NMR) spectroscopy. Protein constructs of different lengths, and with and without a His tag attached at the N-terminus, were studied. The various species showed different overall sensitivities toward urea denaturation with stabilities in the following order: hamster ≤ mouse < rabbit < bovine protein. This order is in agreement with recent circular dichroism (CD) spectroscopic measurements for several species [Khan, M. Q. (2010) Proc. Natl. Acad. Sci. U.S.A.107, 19808-19813] and for the bovine protein presented herein. The [urea] 1/2 values determined by CD spectroscopy parallel those of the most stable residues observed by NMR spectroscopy. Neither the longer constructs containing an additional hydrophobic region nor the His tag influenced the stability of the structured domain of the constructs studied. The effect of the S174N mutation in rabbit PrP C was also investigated. The rank order of the regional stabilities within each protein remained the same for all species. In particular, the residues in the β-sheet region in all four species were more sensitive to urea-induced unfolding than residues in the α2 and α3 helical regions. These observations indicate that the regional specific unfolding pattern is the same for the four mammalian prion proteins studied but militate against the idea that PrP Sc formation is linked with the global stability of PrP C. © 2011 American Chemical Society.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC)
Peer reviewedYes
NPARC number21271637
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Record identifier8e737c2f-4b6f-4de4-81c6-b7fa09445e75
Record created2014-03-24
Record modified2016-05-09
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