Allelic Origin of Protease-Sensitive and Protease-Resistant Prion Protein Isoforms in Gerstmann-Sträussler-Scheinker Disease with the P102L Mutation (original) (raw)
Figure 2
Two-dimensional analysis of mutant and wild-type PrP27-30, C-terminal truncated fragments, and insoluble PrPSc species in GSS P102L.
(A) Immunoblot with 3F4 of PrP27-30 in case # 4 shows three sets of differently glycosylated charge isomers migrating between 30 and 20 kDa, in addition to three spots with a mass of 8 kDa and pIs from 8.4 to 9.6 (arrow). (B) After deglycosylation, the 3F4-reactive core fragment resolves as five spots with net charges between 6.1 and 8.1. (C) ICSM 35 antibody decorates three major spots at 8.1, 7.7, 7.3, and a minor spot at 6.8. (D) Proteinase K-resistant PrPSc probed with 3E2 shows the three sets of PrP27-30, in addition to acidic isoforms migrating at 20 and 14 kDa, which after deglycosylation (E), resolved as a core fragment of six spots at 20 kDa, and C-terminal truncated species of 16–17- and 12–14-kDa. (F) In contrast, the PrP27-30 core fragment associated to E200K mutation resolves as a major spot at pI of 6.8. Immunoblots of the detergent-insoluble fraction, show the presence of naturally occurring PrPSc species with migration overlapping PrP27-30, the 8 kDa fragment (G), and C-terminal truncated fragments (H). (I) Schematic diagram of PK-resistant mutant and wild-type PrPSc forms.