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Positively charged lysine residues, particularly located at codons 24 and 27, are importnat for the pre-OR residues 23–31 to form a PK-resistant structure in prion-infected N2a cells.

By Yoshitaka Yamaguchi (140591), Hironori Miyata (140595), Keiji Uchiyama (140599), Akira Ootsuyama (140602), Sachiko Inubushi (140606), Tsuyoshi Mori (140609), Naomi Muramatsu (140613), Shigeru Katamine (140616) and Suehiro Sakaguchi (140620)


<p>(A) Amino acid sequences of the pre-OR residues 23–31 in mutant proteins. Bold residues indicate substituted residues. (B) Western blotting of N2aC24L1-3 cells transfected with control pcDNA3.1(+) and expression vectors encoding each mutant protein using 3F4 anti-PrP antibodies. The cell lysates were treated with PK at 5 µg/ml. All of the mutant proteins were converted into PK-resistant isoforms in N2aC24L1-3 cells, and all of the mutant isoforms gave rise to doublet non-glycosylated and mono-glycosylated bands. The doublet bands of moPrP(3F4)<sup>Sc</sup>Δ32–88(K23A), moPrP(3F4)<sup>Sc</sup>Δ32–88(K24A) and moPrP(3F4)<sup>Sc</sup>Δ32–88(K27A) were similar in molecular size to those of moPrP(3F4)<sup>Sc</sup>Δ32–88. However, moPrP(3F4)<sup>Sc</sup>Δ32–88(K24,27A) gave rise to the doublet band with the upper band migrating very closely to the lower band, similarly to moPrP(3F4)<sup>Sc</sup>Δ32–88(3K3A). MoPrP(3F4)<sup>Sc</sup>Δ32–88(K23,24A) and moPrP(3F4)<sup>Sc</sup>Δ32–88(K23,27A) showed the upper band with an intermediate molecular size. MoPrP(3F4)<sup>Sc</sup>Δ32–88(3K3R) giving rise to doublet bands with similar molecular size to those of moPrP(3F4)<sup>Sc</sup>Δ32–88. (C) IBL-N antibodies recognized all of the PK-resistant isoforms except for moPrP(3F4)<sup>Sc</sup>Δ32–88(3K3A) and moPrP(3F4)<sup>Sc</sup>Δ32–88(K24,27A).</p

Topics: Infectious Diseases, charged, lysine, located, codons, 24, importnat, pre-or, residues, pk-resistant, prion-infected, n2a
Year: 2012
DOI identifier: 10.1371/journal.pone.0043540.g008
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Provided by: FigShare
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