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Comment
. 2002 Jul 9;99(14):9098-100.
doi: 10.1073/pnas.152318899. Epub 2002 Jul 1.

Progress toward an ultimate proof of the prion hypothesis

Susan W Liebman  1
Affiliations
Comment

Progress toward an ultimate proof of the prion hypothesis

Susan W Liebman. Proc Natl Acad Sci U S A. .
No abstract available

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Figures

Figure 1
Figure 1
Appearance of [Het-s] after microprojectile bombardment of different forms of recombinant HET-s into [Het-s*] prion-free mycelia. Starting with purified His-tagged HET-s isolated from E. coli under denaturing conditions (Upper), Maddelein et al. (16) produced freshly renatured HET-s that was soluble and monomeric. After 72 h, the renatured HET-s spontaneously formed amyloid-like fibers that were separated from the remaining soluble protein by centrifugation. Limited protease K digestion of these fibers resulted in fibers with a 3- to 5-fold reduction in diameter, composed of 7-kDa fragments. Amorphous aggregates were formed by boiling soluble HET-s. Each of these forms of HET-s was layered on mycelia and then bombarded with tungsten particles after evaporation. Bombardment was directly shown to insert fluorescein-labeled undigested HET-s fibers into mycelia. Although the insertion of the other HET-s forms is shown in the cartoon, their actual insertion is an assumption that has not been demonstrated experimentally. Mycelia were scored for the presence of the [Het-s] prion 2 days after the bombardment by testing samples of them for compatibility with [Het-S].
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