Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 20 December 1996:
Vol. 274. no. 5295, pp. 2079 - 2082
DOI: 10.1126/science.274.5295.2079
Prev | Table of Contents | Next

Reports

Evidence for the Conformation of the Pathologic Isoform of the Prion Protein Enciphering and Propagating Prion Diversity

Glenn C. Telling, Piero Parchi, Stephen J. DeArmond, Pietro Cortelli, Pasquale Montagna, Ruth Gabizon, James Mastrianni, Elio Lugaresi, Pierluigi Gambetti, Stanley B. Prusiner *

The fundamental event in prion diseases seems to be a conformational change in cellular prion protein (PrPC) whereby it is converted into the pathologic isoform PrPSc. In fatal familial insomnia (FFI), the protease-resistant fragment of PrPSc after deglycosylation has a size of 19 kilodaltons, whereas that from other inherited and sporadic prion diseases is 21 kilodaltons. Extracts from the brains of FFI patients transmitted disease to transgenic mice expressing a chimeric human-mouse PrP gene about 200 days after inoculation and induced formation of the 19-kilodalton PrPSc fragment, whereas extracts from the brains of familial and sporadic Creutzfeldt-Jakob disease patients produced the 21-kilodalton PrPSc fragment in these mice. The results presented indicate that the conformation of PrPSc functions as a template in directing the formation of nascent PrPSc and suggest a mechanism to explain strains of prions where diversity is encrypted in the conformation of PrPSc.

G. C. Telling and J. Mastrianni, Department of Neurology, University of California, San Francisco, CA 94143, USA.
P. Parchi and P. Gambetti, Division of Neuropathology, Department of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA.
S. J. DeArmond, Departments of Neurology and Pathology, University of California, San Francisco, CA 94143, USA.
P. Cortelli, P. Montagna, E. Lugaresi, Department of Neurology, University of Bologna, Bologna 40123, Italy.
R. Gabizon, Department of Neurology, Hadassah Medical Center, Hebrew University, Jerusalem 91120, Israel.
S. B. Prusiner, Departments of Neurology and Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.
*   To whom correspondence should be addressed at the Department of Neurology, HSE-781, University of California, San Francisco, CA 94143-0518, USA.

For many years the prion diseases, also called transmissible spongiform encephalopathies, were thought to be caused by slow-acting viruses (1), but it is now clear that prions are not viruses and that they are devoid of nucleic acid (2, 3). Prions seem to be composed only of PrPSc molecules, which are abnormal conformers of a normal, host-encoded protein designated PrPC (3, 4). PrPC has a high alpha -helical content and is virtually devoid of beta -sheets, whereas PrPSc has a high beta -sheet content (4, 5); thus, the conversion of PrPC into PrPSc involves a profound conformational change. Formation of PrPSc is a posttranslational process that does not appear to involve a covalent modification of the protein (6).

The prion diseases are unique in that they may present as inherited and infectious disorders (3, 7). More than 20 different mutations of the human (Hu) PrP gene segregate with dominantly inherited disease; five of these have been genetically linked to familial Creutzfeldt-Jakob disease (fCJD), Gerstmann-Sträussler-Scheinker disease, and fatal familial insomnia (FFI) (8). The most common prion diseases of animals are scrapie of sheep and bovine spongiform encephalopathy; the latter may have been transmitted to people through foods (9).

To extend studies on the transmission of wild-type and mutant prions from sporadic Creutzfeldt-Jakob disease (sCJD) and fCJD patients, respectively, to transgenic mice expressing a chimeric mouse-human PrP gene [Tg(MHu2M) mice] (10, 11), we inoculated these mice with mutant prions from the brains of patients who died of FFI. Transmission of human prions to Tg(MHu2M) mice involves the conversion of chimeric MHu2M PrPC into MHu2M PrPSc through a process that is thought to involve the binding of PrPSc to PrPC as PrPC undergoes a structural transition (12, 13). A point mutation of the PrP gene at codon 178 [in which an Asp residue at position 178 is mutated to Asn (D178N)] is the cause of FFI, but a Met residue must be encoded at position 129 on the mutant allele for the FFI phenotype to be manifest (14). The same D178N mutation segregates with a subtype of fCJD, but in this case, Val is encoded on the mutant allele at position 129. The D178N mutation is thought to destabilize the structure of PrPC, resulting in its transformation into PrPSc (13, 15). Some investigators have reported transmission of FFI prions to non-Tg and Tg(HuPrP) mice; the incubation times exceeded 400 days, and only a minority of the inoculated Tg(HuPrP) mice expressing both human and mouse PrPC developed disease (16). These findings with Tg(HuPrP) mice are in accord with earlier studies showing that transmission of human prions to Tg(HuPrP) mice is inhibited by mouse PrPC, and this inhibition can be abolished by ablation of the mouse PrP gene (Prnp0/0) (10, 11).

Tg(MHu2M)Prnp0/0 mice (17) were inoculated intracerebrally with extracts prepared from brain tissue obtained after the death of individuals who died of FFI, fCJD(E200K) (with a mutation in which Glu at position 200 has mutated to Lys), or sCJD. The mice developed signs of experimental prion disease about 200 days after inoculation (Table 1). At the time of writing, inoculation of Tg(MHu2M)Prnp0/0 mice has resulted in primary passage of prions from at least one brain region from three of four FFI patients. As previously reported, Tg(MHu2M)Prnp0/0 mice are susceptible to prions from patients who carried the E200K mutation (11). Extracts from patients who died with fCJD(E200K) or sCJD(M/M129) (homozygous for Met at position 129) caused neurologic dysfunction in Tg(MHu2M)Prnp0/0 mice between 170 and 190 days after inoculation.

Table 1. Transmission of neurodegeneration from patients with FFI or CJD to Tg(MHu2M)Prnp0/0 mice. M, Met; V, Val; n, number with central nervous sytem dysfunction; no, number inoculated.

Patient* Brain region Codon 129  Incubation time mean days ± SEM (n/no)
FFI (IV-37) Frontal cortex M/M 193 ± 5 (9/9) FFI
FFI (IV-26) Thalamus M/M 206 ± 7 (7/7)
FFI (IV-26) Frontal cortex M/M 232 ± 9 (6/6)
FFI (IV-26) Cerebellum M/M >400 (0/10)
FFI (IV-16) Frontal cortex M/V 222 ± 7 (7/7)
FFI (V-58) Frontal cortex M/V >350 (0/10)
FFI (V-58) Insula M/V >350 (0/10)
fCJD (CM, D178N) Parietal cortex V/V >400 (0/10)
sCJD (WL) Frontal cortex M/M 186 ± 5 (9/9)
sCJD (MA)dagger Frontal cortex M/M 180 ± 5 (8/8)
fCJD (LJ1, E200K)dagger Frontal cortex M/M 170 ± 2 (10/10)
fCJD (LJ2, E200K) Frontal cortex M/M 179 ± 1 (8/8)
fCJD (LJ3, E200K) Frontal cortex M/M 184 ± 4 (8/8)
* All samples were 10% (w/v) brain homogenates that were diluted 1:10 before inoculation. If the PrP gene of the patient carried a mutation other than that found in FFI, then the mutation is noted after the patient's initials or identification code.
dagger Transmissions previously reported (11).

The failure to transmit disease with brain homogenates of frontal and insula cortices from FFI patient V-58 is apparently not related to heterozygosity at codon 129, because homogenate from patient IV-16, who has the same haplotype, transmitted the disease. Patients V-58 and IV-16 are phenotypically similar and exemplify the FFI phenotype of the codon 129 heterozygotes with especially long duration (18). The sleep disorder was comparable in patients V-58 and IV-16, and spongiosis was actually more severe in patient IV-16 than in patient V-58 (18). It is noteworthy that a homogenate prepared from the parietal cortex of a patient with fCJD(D178N, V/V129) has so far failed to transmit disease. Whether the titer of prions in this particular sample is low or the Val (V) residue at position 129 in combination with the D178N mutation prevents transmission to Tg(MHu2M)Prnp0/0 mice remains to be established.

Prion proteins in extracts from the brains of Tg(MHu2M) mice inoculated with FFI(D178N, M129) were compared with those inoculated with fCJD(E200K) or sCJD. Mouse brain homogenates were digested with proteinase K (100 µg/ml) for 1 hour at 37°C followed by denaturation by boiling in 3% SDS. The denatured PrPSc was then digested with glycopeptide N- glycosidase (PNGase F) to remove Asn-linked oligosaccharides. As previously described, the human brain extracts prepared from FFI patients yielded a 19-kD protein, whereas extracts from brains of patients with fCJD(E200K) or typical sCJD contained a 21-kD protein (19). Because the amino acid sequences of HuPrPSc molecules from FFI and fCJD(E200K) patients differ at two residues, it was not surprising that the conformations of PrPSc as reflected by the size of the protease-resistant PrP fragments are different (Fig. 1A). In contrast, it was unexpected that PrPSc found in Tg(MHu2M) mice inoculated with FFI prions would be 19 kD, whereas that in Tg(MHu2M) mice injected with fCJD(E200K) was 21 kD (Fig. 1A). These findings demonstrate that the conformation of HuPrPSc in the inoculum is replicated in the brains of the Tg(MHu2M)Prnp0/0 mice by conversion of MHu2M PrPC into MHu2M PrPSc. Wild-type PrPSc in the brain extract from a patient who died of sCJD was found to be 21 kD. Transmission of sCJD to Tg(MHu2M)Prnp0/0 mice produced MHu2M PrPSc, also of size 21 kD, again demonstrating the fidelity of the conversion process whereby the conformation of MHu2M PrPSc in the Tg(MHu2M)Prnp0/0 mouse reflects that of HuPrPSc in the inoculum (Fig. 1B). We emphasize that whereas the primary structures of the PrPSc molecules in the three different human brain inocula are distinct, the amino acid sequences of the PrPSc molecules in the brains of inoculated Tg(MHu2M) mice are invariant. The MHu2M PrP transgene was sequenced and found to be the same as the construct used for microinjections during production of the mice (20).

Fig. 1. Protein immunoblot analysis of PrPSc from brains of Tg(MHu2M) mice inoculated with FFI, fCJD(E200K), and sCJD. Homogenates of human or mouse brain were prepared as described (30). (A) Comparison of fCJD(E200K) and FFI. Samples analyzed are from the following sources: lane 1, fCJD(E200K) patient LJ1, frontal cortex; lane 2, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used for lane 1; lane 3, fCJD(E200K) patient LJ1, frontal cortex treated with PNGase F; lane 4, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used in lane 3; lane 5, FFI patient IV-16, frontal cortex; lane 6, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used for lane 5; lane 7, FFI patient IV-16, frontal cortex treated with PNGase F; and lane 8, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used in lane 7. (B) Comparison of sCJD and FFI. Samples analyzed are from the following sources: lane 1, sCJD patient EC; lane 2, Tg(MHu2M) mouse inoculated with preparation used in lane 1; lane 3, sCJD patient EC treated with PNGase F; lane 4, Tg(MHu2M) mouse inoculated with preparation used in lane 3; lane 5, FFI patient IV-16, frontal cortex; lane 6, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used in lane 5; lane 7, FFI patient IV-16, frontal cortex treated with PNGase F; lane 8, Tg(MHu2M)Prnp0/0 mouse inoculated with preparation used in lane 7. [View Larger Version of this Image (62K GIF file)]

We also examined the regional distribution of PrPSc in the brains of Tg(MHu2M)Prnp0/0 mice inoculated with prions from FFI patients as well as from fCJD(E200K) and sCJD patients (21). Histoblots of coronal brain sections through the hippocampus and thalamus as well as those through the brainstem and cerebellum were developed (Fig. 2). The pattern of PrPSc deposition in FFI-inoculated Tg(MHu2M)Prnp0/0 mice was clearly different from Tg(MHu2M)Prnp0/0 mice inoculated with CJD.

Fig. 2. Regional distribution of PrPSc in Tg(MHu2M)Prnp0/0 mice inoculated with extracts from FFI, CJD(E200K), and sCJD patients. Histoblots of coronal sections of the brain and brainstem of mice inoculated with extracts from FFI (A and E), fCJD(E200K) (B and F), or sCJD(M/M129) (C and G) patients were performed as described (21). PrPC was eliminated from the section by exposing the membranes for 18 hours at 37°C to proteinase K (400 µg/ml) in a buffer containing 0.5% Brij 35, 100 mM NaCl, and 10 mM tris-HCl, pH 7.8. Immunostaining of PrPSc was enhanced by exposing the histoblots to 3 M guanidinium isothiocyanate for 10 min at room temperature in 20 mM tris-HCl, pH 7.8, then rinsing three times with TBST [10 mM tris-HCl, pH 8.0, 150 mM NaCl, 0.5% (v/v) Tween 20] before immunostaining with monoclonal antibody 3F4 to PrP (anti-PrP) (31). Also shown are labeled diagrams of the coronal sections of the hippocampus-thalamus region (D) and brainstem-cerebellum region (H). NC, neocortex; Hp, hippocampus; Hb, habenula; Th, thalamus; vpl, ventral posterior lateral thalamic nucleus; Hy, hypothalamus; Am, amygdala; GC, granular cell layer of the cerebellum; IC, inferior colliculus; R, dorsal nucleus of the raphe; LC, locus ceruleus. [View Larger Version of this Image (89K GIF file)]

In FFI-inoculated mice, PrPSc deposition was most intense in the thalamus and the rostral part of the corpus callosum (Fig. 2A). In FFI patients PrPSc deposition and neuropathologic changes are marked in the antero-ventral and mediodorsal nuclei of the thalamus (18, 22). Intermediate intensities of immunostaining were found in the deeper layers of the frontal cortex and in the lateral portions of the caudate nuclei. Little or no immunostaining was found in the habenula or the hypothalamus (Fig. 2A). Staining in the hippocampus was also negative except for the stratum lacunosum molecularae where most of the spongiform degeneration and reactive astrocytic gliosis occurred. The absence of PrPSc deposition in the habenula is unique to FFI because deposition invariably occurs in this region in response to CJD and scrapie prion inoculations (10, 23).

In contrast to FFI-inoculated mice, inoculation of Tg(MHu2M) mice with fCJD(E200K) and sCJD prions induced PrPSc accumulation in many areas of the central nervous system (Fig. 2, B and C). Although inoculation with fCJD(E200K) and sCJD as well as with iatrogenic CJD prions (24) resulted in accumulations of PrPSc in the brainstem (Fig. 2, F and G), that was not the case for FFI (Fig. 2E). These differences in PrPSc deposition reflect earlier studies on prion strains where the patterns of spongiform degeneration and PrPSc accumulation were specific for a particular strain when assessed in isogenic animals (23, 25).

The neuropathologic changes in the brains of five Tg(MHu2M)Prnp0/0 mice inoculated with prions from three different FFI patients were examined (Fig. 3). They were characterized by moderate to severe spongiform degeneration and astrocytic gliosis in the deeper layers of the frontal cortex and rostral part of the cingulate gyrus, the thalamus, the lateral portions of the caudate nucleus, and in the white matter tracts of the cerebral hemispheres. Immunohistochemical examination of FFI-inoculated brains (Fig. 3B) showed that regions with the largest amount of PrP staining corresponded to the regions with the most severe neuropathological changes in both gray and white matter. The accentuated immunostaining resulted from multiple primitive PrP plaques ranging in size from 10 to 40 µM (Fig. 3B).

Fig. 3. Representative examples of neuropathologic changes in Tg(MHu2M)Prnp0/0 mice after inoculation with human FFI prions. (A) Hematoxylin and eosin stain of a serial section of the thalamus shows mild to moderate spongiform degeneration of the neuropil, with vacuoles 10 to 30 µm in diameter. Brain tissue was immersion-fixed in 10% buffered formalin solution after the animals were killed. The brains were embedded in paraffin and histological sections prepared and stained with hematoxylin and eosin for evaluation of spongiform degeneration. (B) PrP immunohistochemistry of a serial section of the thalamus shows multiple punctate PrP-immunopositive deposits, the largest being sim 10 µm. PrP immunoreactivity was enhanced by immersing the sections in 1.3 mM HCl and autoclaving them at 121°C for 10 min (32). Immunostaining of tissue sections was performed as previously described (33) with anti-PrP (31). Bar in (B) = 50 µm and also applies to (A). [View Larger Version of this Image (124K GIF file)]

Comparison of FFI-inoculated Tg- (MHu2M)Prnp0/0 mice with the same type mice inoculated with prions from fCJD- (E200K) and sCJD patients revealed two main histopathologic differences. First, FFI produced no vacuolation in the hypothalamus, whereas a mild to moderate degree of vacuolation was found with both sCJD and fCJD(E200K) prions. Secondly, FFI produced moderate to severe vacuolation of the corpus callosum. In contrast, there was only mild vacuolation of the corpus callosum with sCJD and none with fCJD(E200K).

Our studies show that human prions inoculated into Tg(MHu2M)Prnp0/0 mice instruct with substantial fidelity the formation of distinct MHu2M PrPSc molecules. Although the prions in patients with FFI, fCJD(E200K), and sCJD can be distinguished by differences in the amino acid sequence of HuPrPSc, that is clearly not the case for MHu2M PrPSc in the brains of the inoculated Tg(MHu2M)Prnp0/0 mice. From our findings we contend that the different sizes of chimeric MHu2M PrPSc molecules found after limited digestion with proteinase K result from distinct secondary and tertiary structures. Whether these differences in PrPSc structure will be propagated upon serial passage of the chimeric prions in Tg(MHu2M)Prnp0/0 or Tg(HuPrP)Prnp0/0 mice remains to be determined.

If such properties are propagated, then this will suggest that different mutant human PrPs will have generated distinct strains of prions. The existence of prion strains has posed a conundrum as to the mechanism by which strain-specific characteristics are encrypted (23, 26). Although differences in the size of protease-resistant fragments of PrPSc have not been a general characteristic of prion strains (27), the hyper and drowsy strains of prions isolated from mink by serial passage in Syrian hamsters do differ with respect to the size of the PrPSc molecules after limited proteolysis (28). But unlike the studies reported here, where prions were generated de novo in patients carrying the D178N or E200K mutations, the origin of the hyper and drowsy strains is obscure.

Our results provide a plausible mechanism for explaining diversity in a pathogen that lacks nucleic acid; the biological properties of prion strains seem to be encrypted in the conformation of PrPSc. Because prion strains produce different disease phenotypes, such findings raise the possibility that deviations in the phenotypes of other degenerative disorders may also reflect conformational variants in pathologic proteins. Variations in the conformation of PrPSc are reproduced through templating of the PrPSc in the inoculum onto the substrate PrPC. Deciphering the molecular events by which the conformation of one protein is imparted to another and the mechanism responsible for the apparently high degree of fidelity associated with this process should be of considerable interest. Indeed, the foregoing data violate the widely and long-held idea that amino acid sequences are the sole determinants of the tertiary structures of biologically active proteins (29).

REFERENCES AND NOTES

  1. D. C. Gajdusek, Science 197, 943 (1977) [Medline].
  2. S. B. Prusiner, ibid. 216, 136 (1982) [Medline].
  3. ___, ibid. 252, 1515 (1991) [Medline].
  4. K.-M. Pan et al., Proc. Natl. Acad. Sci. U.S.A. 90, 10962 (1993) [Medline].
  5. S. B. Prusiner et al., Cell 35, 349 (1983) [Medline]; B. W. Caughey et al., Biochemistry 30, 7672 (1991) [Medline]; J. Safar, P. P. Roller, D. C. Gajdusek, C. J. Gibbs Jr., J. Biol. Chem. 268, 20276 (1993) [Medline]; P. Pergami, H. Jaffe, J. Safar, Anal. Biochem. 236, 63 (1996) [Medline].
  6. D. R. Borchelt, M. Scott, A. Taraboulos, N. Stahl, S. B. Prusiner, J. Cell Biol. 110, 743 (1990) [Medline]; N. Stahl et al., Biochemistry 32, 1991 (1993) [Medline].
  7. F. Meggendorfer, Z. Gesmate. Neurol. Psychiatr. 128, 337 (1930); C. L. Masters, D. C. Gajdusek, C. J. Gibbs Jr., Brain 104, 559 (1981) [Medline].
  8. K. Hsiao et al., Nature 338, 342 (1989) [Medline]; M. Poulter et al., Brain 115, 675 (1992) [Medline]; S. R. Dlouhy et al., Nature Genet. 1, 64 (1992) [Medline]; R. B. Petersen et al., Neurology 42, 1859 (1992) [Medline]; R. Gabizon et al., Am. J. Hum. Genet. 33, 828 (1993) .
  9. R. M. Anderson et al., Nature 382, 779 (1996) [Medline]; G. Chazot et al., Lancet 347, 1181 (1996) [Medline]; R. G. Will et al., ibid., p. 921.
  10. G. C. Telling et al., Proc. Natl. Acad. Sci. U.S.A. 91, 9936 (1994) [Medline].
  11. G. C. Telling et al., Cell 83, 79 (1995) [Medline].
  12. S. B. Prusiner et al., ibid. 63, 673 (1990) [Medline].
  13. F. E. Cohen et al., Science 264, 530 (1994) [Medline].
  14. R. Medori et al., N. Engl. J. Med. 326, 444 (1992) [Medline]; L. G. Goldfarb et al., Science 258, 806 (1992) [Medline].
  15. R. B. Petersen, P. Parchi, S. L. Richardson, C. B. Urig, P. Gambetti, J. Biol. Chem. 271, 122661 (1996) ; R. Riek et al., Nature 382, 180 (1996) [Medline].
  16. J. Tateishi et al., Nature 376, 434 (1995) [Medline]; J. Collinge et al., Lancet 346, 569 (1995) [Medline].
  17. In contrast to Tg(HuPrP) mice, Tg(MHu2M) mice are susceptible to human prions (10). When Tg(MHu2M)Prnp0/0 mice were produced by crossing onto the PrP null background, a 20% reduction in incubation times was found (11).
  18. P. Parchi et al., Ann. Neurol. 38, 21 (1995) [Medline].
  19. L. Monari et al., Proc. Natl. Acad. Sci. U.S.A. 91, 2839 (1994) [Medline]; P. Parchi et al., Ann. Neurol. 39, 767 (1996) [Medline].
  20. G. Telling, N. Heye, S. B. Prusiner, unpublished data.
  21. A. Taraboulos et al., Proc. Natl. Acad. Sci. U.S.A. 89, 7620 (1992) [Medline].
  22. V. Manetto et al., Neurology 42, 312 (1992) [Medline]; P. Gambetti, P. Parchi, R. B. Petersen, S. G. Chen, E. Lugaresi, Brain Pathol. 5, 43 (1995) [Medline].
  23. R. Hecker et al., Genes Dev. 6, 1213 (1992) [Medline].
  24. S. J. DeArmond, G. C. Telling, S. B. Prusiner, unpublished data.
  25. H. Fraser and A. G. Dickinson, J. Comp. Pathol. 83, 29 (1973) [Medline]; M. E. Bruce, A. G. Dickinson, H. Fraser, Neuropathol. Appl. Neurobiol. 2, 471 (1976) ; S. J. DeArmond et al., Proc. Natl. Acad. Sci. U.S.A. 90, 6449 (1993).
  26. A. G. Dickinson, V. M. H. Meikle, H. Fraser, J. Comp. Pathol. 78, 293 (1968) [Medline]; M. E. Bruce and A. G. Dickinson, J. Gen. Virol. 68, 79 (1987) [Medline]; R. H. Kimberlin, C. A. Walker, H. Fraser, ibid. 70, 2017 (1989) [Medline]; R. I. Carp and S. M. Callahan, ibid. 72, 293 (1991).
  27. M. Scott, S. J. DeArmond, S. B. Prusiner, unpublished data.
  28. R. A. Bessen and R. F. Marsh, J. Gen. Virol. 73, 329 (1992) [Medline]; J. Virol. 68, 7859 (1994).
  29. C. B. Anfinsen, Science 181, 223 (1973) [Medline].
  30. Homogenates (10%, w/v) of human or mouse brain were prepared by repeated extrusion through an 18-gauge syringe needle followed by a 22-gauge needle in phosphate-buffered saline lacking calcium and magnesium ions. For immunoblot analysis, samples were adjusted to 0.5% NP-40 and 0.5% sodium deoxycholate, and samples were digested with proteinase K (PK) (100 µg/ml) for 1 hour at 37°C. Digestion was terminated by the addition of phenylmethylsufonylfluoride (2 mM final concentration) and boiling in electrophoresis sample buffer (3% SDS, 62.5 mM tris, pH 6.8). For deglycosylation, the PK-treated samples were digested for 2 hours with recombinant PNGase F (New England Biolabs) as specified by the supplier, precipitated with four volumes of methanol at -20°C, and resuspended in electrophoresis sample buffer.
  31. R. J. Kascsak et al., J. Virol. 61, 3688 (1987) [Medline].
  32. T. Muramoto, T. Kitamoto, J. Tateishi, I. Goto, Am. J. Pathol. 140, 1411 (1992) [Medline].
  33. G. C. Telling et al., Genes Dev. 10, 1736 (1996) [Medline].
  34. This work was supported by grants from NIH and the American Health Assistance Foundation, as well as by gifts from the Sherman Fairchild Foundation and the Britton Fund. G.T. was supported by a fellowship from an NIH postdoctoral training grant (NS07219).

23 August 1996; accepted 28 October 1996


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Absence of spontaneous disease and comparative prion susceptibility of transgenic mice expressing mutant human prion proteins.
E. A. Asante, I. Gowland, A. Grimshaw, J. M. Linehan, M. Smidak, R. Houghton, O. Osiguwa, A. Tomlinson, S. Joiner, S. Brandner, et al. (2009)
J. Gen. Virol. 90, 546-558
   Abstract »    Full Text »    PDF »
From the Cover: The yeast Sup35NM domain propagates as a prion in mammalian cells.
C. Krammer, D. Kryndushkin, M. H. Suhre, E. Kremmer, A. Hofmann, A. Pfeifer, T. Scheibel, R. B. Wickner, H. M. Schatzl, and I. Vorberg (2009)
PNAS 106, 462-467
   Abstract »    Full Text »    PDF »
Prion Strain Targeting Independent of Strain-Specific Neuronal Tropism.
J. I. Ayers, A. E. Kincaid, and J. C. Bartz (2009)
J. Virol. 83, 81-87
   Abstract »    Full Text »    PDF »
Specific Biarsenical Labeling of Cell Surface Proteins Allows Fluorescent- and Biotin-tagging of Amyloid Precursor Protein and Prion Proteins.
Y. Taguchi, Z.-D. Shi, B. Ruddy, D. W. Dorward, L. Greene, and G. S. Baron (2009)
Mol. Biol. Cell 20, 233-244
   Abstract »    Full Text »    PDF »
The bank vole (Myodes glareolus) as a sensitive bioassay for sheep scrapie.
M. A. Di Bari, F. Chianini, G. Vaccari, E. Esposito, M. Conte, S. L. Eaton, S. Hamilton, J. Finlayson, P. J. Steele, M. P. Dagleish, et al. (2008)
J. Gen. Virol. 89, 2975-2985
   Abstract »    Full Text »    PDF »
The origin of the prion agent of kuru: molecular and biological strain typing.
J. D.F Wadsworth, S. Joiner, J. M Linehan, E. A Asante, S. Brandner, and J. Collinge (2008)
Phil Trans R Soc B 363, 3747-3753
   Abstract »    Full Text »    PDF »
Molecular structural basis for polymorphism in Alzheimer's {beta}-amyloid fibrils.
A. K. Paravastu, R. D. Leapman, W.-M. Yau, and R. Tycko (2008)
PNAS 105, 18349-18354
   Abstract »    Full Text »    PDF »
Molecular and Transmission Characteristics of Primary-Passaged Ovine Scrapie Isolates in Conventional and Ovine PrP Transgenic Mice.
A. M. Thackray, L. Hopkins, J. Spiropoulos, and R. Bujdoso (2008)
J. Virol. 82, 11197-11207
   Abstract »    Full Text »    PDF »
Characterization of Truncated Forms of Abnormal Prion Protein in Creutzfeldt-Jakob Disease.
S. Notari, R. Strammiello, S. Capellari, A. Giese, M. Cescatti, J. Grassi, B. Ghetti, J. P. M. Langeveld, W.-Q. Zou, P. Gambetti, et al. (2008)
J. Biol. Chem. 283, 30557-30565
   Abstract »    Full Text »    PDF »
A novel PRNP-P105S mutation associated with atypical prion disease and a rare PrPSc conformation.
E. Tunnell, R. Wollman, S. Mallik, C. J. Cortes, S. J. DeArmond, and J. A. Mastrianni (2008)
Neurology 71, 1431-1438
   Abstract »    Full Text »    PDF »
The effects of prion protein proteolysis and disaggregation on the strain properties of hamster scrapie.
A. M. Deleault, N. R. Deleault, B. T. Harris, J. R. Rees, and S. Supattapone (2008)
J. Gen. Virol. 89, 2642-2650
   Abstract »    Full Text »    PDF »
The Same Primary Structure of the Prion Protein Yields Two Distinct Self-propagating States.
N. Makarava and I. V. Baskakov (2008)
J. Biol. Chem. 283, 15988-15996
   Abstract »    Full Text »    PDF »
Kuru prions and sporadic Creutzfeldt-Jakob disease prions have equivalent transmission properties in transgenic and wild-type mice.
J. D. F. Wadsworth, S. Joiner, J. M. Linehan, M. Desbruslais, K. Fox, S. Cooper, S. Cronier, E. A. Asante, S. Mead, S. Brandner, et al. (2008)
PNAS 105, 3885-3890
   Abstract »    Full Text »    PDF »
The elk PRNP codon 132 polymorphism controls cervid and scrapie prion propagation.
K. M. Green, S. R. Browning, T. S. Seward, J. E. Jewell, D. L. Ross, M. A. Green, E. S. Williams, E. A. Hoover, and G. C. Telling (2008)
J. Gen. Virol. 89, 598-608
   Abstract »    Full Text »    PDF »
Mouse-Adapted Ovine Scrapie Prion Strains Are Characterized by Different Conformers of PrPSc.
A. M. Thackray, L. Hopkins, M. A. Klein, and R. Bujdoso (2007)
J. Virol. 81, 12119-12127
   Abstract »    Full Text »    PDF »
A General Model of Prion Strains and Their Pathogenicity.
J. Collinge and A. R. Clarke (2007)
Science 318, 930-936
   Abstract »    Full Text »    PDF »
Molecular Behaviors of "CH1641-Like" Sheep Scrapie Isolates in Ovine Transgenic Mice (TgOvPrP4).
T. Baron and A.-G. Biacabe (2007)
J. Virol. 81, 7230-7237
   Abstract »    Full Text »    PDF »
Molecular Discrimination of Atypical Bovine Spongiform Encephalopathy Strains from a Geographical Region Spanning a Wide Area in Europe.
J. G. Jacobs, J. P. M. Langeveld, A.-G. Biacabe, P.-L. Acutis, M. P. Polak, D. Gavier-Widen, A. Buschmann, M. Caramelli, C. Casalone, M. Mazza, et al. (2007)
J. Clin. Microbiol. 45, 1821-1829
   Abstract »    Full Text »    PDF »
Continuum of prion protein structures enciphers a multitude of prion isolate-specified phenotypes.
G. Legname, H.-O. B. Nguyen, D. Peretz, F. E. Cohen, S. J. DeArmond, and S. B. Prusiner (2006)
PNAS 103, 19105-19110
   Abstract »    Full Text »    PDF »
A systematic review of prion therapeutics in experimental models.
C. R Trevitt and J. Collinge (2006)
Brain 129, 2241-2265
   Abstract »    Full Text »    PDF »
Prion strain-dependent differences in conversion of mutant prion proteins in cell culture..
R. Atarashi, V. L. Sim, N. Nishida, B. Caughey, and S. Katamine (2006)
J. Virol. 80, 7854-7862
   Abstract »    Full Text »    PDF »
Dissociation of pathological and molecular phenotype of variant Creutzfeldt-Jakob disease in transgenic human prion protein 129 heterozygous mice.
E. A. Asante, J. M. Linehan, I. Gowland, S. Joiner, K. Fox, S. Cooper, O. Osiguwa, M. Gorry, J. Welch, R. Houghton, et al. (2006)
PNAS 103, 10759-10764
   Abstract »    Full Text »    PDF »
Probing the Conformation of the Prion Protein within a Single Amyloid Fibril Using a Novel Immunoconformational Assay.
V. Novitskaya, N. Makarava, A. Bellon, O. V. Bocharova, I. B. Bronstein, R. A. Williamson, and I. V. Baskakov (2006)
J. Biol. Chem. 281, 15536-15545
   Abstract »    Full Text »    PDF »
Phenotypic heterogeneity in inherited prion disease (P102L) is associated with differential propagation of protease-resistant wild-type and mutant prion protein.
J. D. F. Wadsworth, S. Joiner, J. M. Linehan, S. Cooper, C. Powell, G. Mallinson, J. Buckell, I. Gowland, E. A. Asante, H. Budka, et al. (2006)
Brain 129, 1557-1569
   Abstract »    Full Text »    PDF »
Prion infection of mouse neurospheres.
R. K. Giri, R. Young, R. Pitstick, S. J. DeArmond, S. B. Prusiner, and G. A. Carlson (2006)
PNAS 103, 3875-3880
   Abstract »    Full Text »    PDF »
Inactivation of Prions by Acidic Sodium Dodecyl Sulfate.
D. Peretz, S. Supattapone, K. Giles, J. Vergara, Y. Freyman, P. Lessard, J. G. Safar, D. V. Glidden, C. McCulloch, H.-O. B. Nguyen, et al. (2006)
J. Virol. 80, 322-331
   Abstract »    Full Text »    PDF »
Breaking an Absolute Species Barrier: Transgenic Mice Expressing the Mink PrP Gene Are Susceptible to Transmissible Mink Encephalopathy.
O. Windl, M. Buchholz, A. Neubauer, W. Schulz-Schaeffer, M. Groschup, S. Walter, S. Arendt, M. Neumann, A. K. Voss, and H. A. Kretzschmar (2005)
J. Virol. 79, 14971-14975
   Abstract »    Full Text »    PDF »
Creutzfeldt-Jakob Disease (CJD) with a Mutation at Codon 148 of Prion Protein Gene: Relationship with Sporadic CJD.
M. Pastore, S. S. Chin, K. L. Bell, Z. Dong, Q. Yang, L. Yang, J. Yuan, S. G. Chen, P. Gambetti, and W.-Q. Zou (2005)
Am. J. Pathol. 167, 1729-1738
   Abstract »    Full Text »    PDF »
A newly identified type of scrapie agent can naturally infect sheep with resistant PrP genotypes.
A. Le Dur, V. Beringue, O. Andreoletti, F. Reine, T. L. Lai, T. Baron, B. Bratberg, J.-L. Vilotte, P. Sarradin, S. L. Benestad, et al. (2005)
PNAS 102, 16031-16036
   Abstract »    Full Text »    PDF »
PrP glycoforms are associated in a strain-specific ratio in native PrPSc.
A. Khalili-Shirazi, L. Summers, J. Linehan, G. Mallinson, D. Anstee, S. Hawke, G. S. Jackson, and J. Collinge (2005)
J. Gen. Virol. 86, 2635-2644
   Abstract »    Full Text »    PDF »
Search for a Prion-Specific Nucleic Acid.
J. G. Safar, K. Kellings, A. Serban, D. Groth, J. E. Cleaver, S. B. Prusiner, and D. Riesner (2005)
J. Virol. 79, 10796-10806
   Abstract »    Full Text »    PDF »
Molecular neurology of prion disease.
J Collinge (2005)
J. Neurol. Neurosurg. Psychiatry 76, 906-919
   Abstract »    Full Text »    PDF »
Biological and Biochemical Characteristics of Prion Strains Conserved in Persistently Infected Cell Cultures.
K. Arima, N. Nishida, S. Sakaguchi, K. Shigematsu, R. Atarashi, N. Yamaguchi, D. Yoshikawa, J. Yoon, K. Watanabe, N. Kobayashi, et al. (2005)
J. Virol. 79, 7104-7112
   Abstract »    Full Text »    PDF »
Prion biology relevant to bovine spongiform encephalopathy.
J. Novakofski, M. S. Brewer, N. Mateus-Pinilla, J. Killefer, and R. H. McCusker (2005)
J Anim Sci 83, 1455-1476
   Abstract »    Full Text »    PDF »
Transmission Barriers for Bovine, Ovine, and Human Prions in Transgenic Mice.
M. R. Scott, D. Peretz, H.-O. B. Nguyen, S. J. DeArmond, and S. B. Prusiner (2005)
J. Virol. 79, 5259-5271
   Abstract »    Full Text »    PDF »
Notch-1 activation and dendritic atrophy in prion disease.
N. Ishikura, J. L. Clever, E. Bouzamondo-Bernstein, E. Samayoa, S. B. Prusiner, E. J. Huang, and S. J. DeArmond (2005)
PNAS 102, 886-891
   Abstract »    Full Text »    PDF »
Self-Propagating, Molecular-Level Polymorphism in Alzheimer's {beta}-Amyloid Fibrils.
A. T. Petkova, R. D. Leapman, Z. Guo, W.-M. Yau, M. P. Mattson, and R. Tycko (2005)
Science 307, 262-265
   Abstract »    Full Text »    PDF »
Transmission of Prions from Mule Deer and Elk with Chronic Wasting Disease to Transgenic Mice Expressing Cervid PrP.
S. R. Browning, G. L. Mason, T. Seward, M. Green, G. A. J. Eliason, C. Mathiason, M. W. Miller, E. S. Williams, E. Hoover, and G. C. Telling (2004)
J. Virol. 78, 13345-13350
   Abstract »    Full Text »    PDF »
Characterization of two distinct prion strains derived from bovine spongiform encephalopathy transmissions to inbred mice.
S. E. Lloyd, J. M. Linehan, M. Desbruslais, S. Joiner, J. Buckell, S. Brandner, J. D. F. Wadsworth, and J. Collinge (2004)
J. Gen. Virol. 85, 2471-2478
   Abstract »    Full Text »    PDF »
Synthetic Mammalian Prions.
G. Legname, I. V. Baskakov, H.-O. B. Nguyen, D. Riesner, F. E. Cohen, S. J. DeArmond, and S. B. Prusiner (2004)
Science 305, 673-676
   Abstract »    Full Text »    PDF »
Molecular Analysis of the Protease-Resistant Prion Protein in Scrapie and Bovine Spongiform Encephalopathy Transmitted to Ovine Transgenic and Wild-Type Mice.
T. Baron, C. Crozet, A.-G. Biacabe, S. Philippe, J. Verchere, A. Bencsik, J.-Y. Madec, D. Calavas, and J. Samarut (2004)
J. Virol. 78, 6243-6251
   Abstract »    Full Text »    PDF »
Effects of Different Experimental Conditions on the PrPSc Core Generated by Protease Digestion: IMPLICATIONS FOR STRAIN TYPING AND MOLECULAR CLASSIFICATION OF CJD.
S. Notari, S. Capellari, A. Giese, I. Westner, A. Baruzzi, B. Ghetti, P. Gambetti, H. A. Kretzschmar, and P. Parchi (2004)
J. Biol. Chem. 279, 16797-16804
   Abstract »    Full Text »    PDF »
Flexible N-terminal Region of Prion Protein Influences Conformation of Protease-resistant Prion Protein Isoforms Associated with Cross-species Scrapie Infection in Vivo and in Vitro.
V. A. Lawson, S. A. Priola, K. Meade-White, M. Lawson, and B. Chesebro (2004)
J. Biol. Chem. 279, 13689-13695
   Abstract »    Full Text »    PDF »
From conversion to aggregation: Protofibril formation of the prion protein.
M. L. DeMarco and V. Daggett (2004)
PNAS 101, 2293-2298
   Abstract »    Full Text »    PDF »
Antibody to DNA detects scrapie but not normal prion protein.
W.-Q. Zou, J. Zheng, D. M. Gray, P. Gambetti, and S. G. Chen (2004)
PNAS 101, 1380-1385
   Abstract »    Full Text »    PDF »
Peripheral Tissue Involvement in Sporadic, Iatrogenic, and Variant Creutzfeldt-Jakob Disease: An Immunohistochemical, Quantitative, and Biochemical Study.
M. W. Head, D. Ritchie, N. Smith, V. McLoughlin, W. Nailon, S. Samad, S. Masson, M. Bishop, L. McCardle, and J. W. Ironside (2004)
Am. J. Pathol. 164, 143-153
   Abstract »    Full Text »    PDF »
Destabilizing Interactions Among [PSI+] and [PIN+] Yeast Prion Variants.
M. E. Bradley and S. W. Liebman (2003)
Genetics 165, 1675-1685
   Abstract »    Full Text »    PDF »
Humanized Knock-In Mice Expressing Chimeric Prion Protein Showed Varied Susceptibility to Different Human Prions.
Y. Taguchi, S. Mohri, J. W. Ironside, T. Muramoto, and T. Kitamoto (2003)
Am. J. Pathol. 163, 2585-2593
   Abstract »    Full Text »    PDF »
Identification of Novel Proteinase K-resistant C-terminal Fragments of PrP in Creutzfeldt-Jakob Disease.
W.-Q. Zou, S. Capellari, P. Parchi, M.-S. Sy, P. Gambetti, and S. G. Chen (2003)
J. Biol. Chem. 278, 40429-40436
   Abstract »    Full Text »    PDF »
Regional heterogeneity of cellular prion protein isoforms in the mouse brain.
V. Beringue, G. Mallinson, M. Kaisar, M. Tayebi, Z. Sattar, G. Jackson, D. Anstee, J. Collinge, and S. Hawke (2003)
Brain 126, 2065-2073
   Abstract »    Full Text »    PDF »
Molecular Distinction between Pathogenic and Infectious Properties of the Prion Protein.
R. Chiesa, P. Piccardo, E. Quaglio, B. Drisaldi, S. L. Si-Hoe, M. Takao, B. Ghetti, and D. A. Harris (2003)
J. Virol. 77, 7611-7622
   Abstract »    Full Text »    PDF »
Activation by prion peptide PrP106-126 induces a NF-{kappa}B-driven proinflammatory response in human monocyte-derived dendritic cells.
S. M. Bacot, P. Lenz, M. R. Frazier-Jessen, and G. M. Feldman (2003)
J. Leukoc. Biol. 74, 118-125
   Abstract »    Full Text »    PDF »
Introduction to the transmissible spongiform encephalopathies or prion diseases.
B. Chesebro (2003)
Br. Med. Bull. 66, 1-20
   Abstract »    Full Text »    PDF »
TSE strain variation: An investigation into prion disease diversity.
M. E Bruce (2003)
Br. Med. Bull. 66, 99-108
   Abstract »    Full Text »    PDF »
Sporadic and familial CJD: classification and characterisation.
P. Gambetti, Q. Kong, W. Zou, P. Parchi, and S. G Chen (2003)
Br. Med. Bull. 66, 213-239
   Abstract »    Full Text »    PDF »
Molecular and clinical classification of human prion disease.
J. D. Wadsworth, A. F Hill, J. A Beck, and J. Collinge (2003)
Br. Med. Bull. 66, 241-254
   Abstract »    Full Text »    PDF »
Molecular classification of sporadic Creutzfeldt-Jakob disease.
A. F. Hill, S. Joiner, J. D. F. Wadsworth, K. C. L. Sidle, J. E. Bell, H. Budka, J. W. Ironside, and J. Collinge (2003)
Brain 126, 1333-1346
   Abstract »    Full Text »    PDF »
Distinct profiles of PrPd immunoreactivity in the brain of scrapie- and BSE-infected sheep: implications for differential cell targeting and PrP processing.
L. Gonzalez, S. Martin, and M. Jeffrey (2003)
J. Gen. Virol. 84, 1339-1350
   Abstract »    Full Text »    PDF »
Abbreviated incubation times for human prions in mice expressing a chimeric mouse-human prion protein transgene.
C. Korth, K. Kaneko, D. Groth, N. Heye, G. Telling, J. Mastrianni, P. Parchi, P. Gambetti, R. Will, J. Ironside, et al. (2003)
PNAS 100, 4784-4789
   Abstract »    Full Text »    PDF »
Cellular Prion Protein Sensitizes Neurons to Apoptotic Stimuli through Mdm2-regulated and p53-dependent Caspase 3-like Activation.
E. Paitel, R. Fahraeus, and F. Checler (2003)
J. Biol. Chem. 278, 10061-10066
   Abstract »    Full Text »    PDF »
Microglia from Creutzfeldt-Jakob Disease-Infected Brains Are Infectious and Show Specific mRNA Activation Profiles.
C. A. Baker, D. Martin, and L. Manuelidis (2002)
J. Virol. 76, 10905-10913
   Abstract »    Full Text »    PDF »
Biological and Biochemical Characterization of Sheep Scrapie in Japan.
M. Horiuchi, T. Nemoto, N. Ishiguro, H. Furuoka, S. Mohri, and M. Shinagawa (2002)
J. Clin. Microbiol. 40, 3421-3426
   Abstract »    Full Text »    PDF »
Characterization of Thermodynamic Diversity between Transmissible Spongiform Encephalopathy Agent Strains and Its Theoretical Implications.
R. A. Somerville, R. C. Oberthur, U. Havekost, F. MacDonald, D. M. Taylor, and A. G. Dickinson (2002)
J. Biol. Chem. 277, 11084-11089
   Abstract »    Full Text »    PDF »
Structural studies of the scrapie prion protein by electron crystallography.
H. Wille, M. D. Michelitsch, V. Guenebaut, S. Supattapone, A. Serban, F. E. Cohen, D. A. Agard, and S. B. Prusiner (2002)
PNAS 99, 3563-3568
   Abstract »    Full Text »    PDF »
Inherited prion disease caused by the V210I mutation: Transmission to transgenic mice.
J. A. Mastrianni, S. Capellari, G. C. Telling, D. Han, P. Bosque, S. B. Prusiner, and S. J. DeArmond (2001)
Neurology 57, 2198-2205
   Abstract »    Full Text »    PDF »
The molecular pathology of CJD: old and new variants.
G S Jackson and J Collinge (2001)
Mol. Pathol. 54, 393-399
   Abstract »    Full Text »    PDF »
Long-Term Subclinical Carrier State Precedes Scrapie Replication and Adaptation in a Resistant Species: Analogies to Bovine Spongiform Encephalopathy and Variant Creutzfeldt-Jakob Disease in Humans.
R. Race, A. Raines, G. J. Raymond, B. Caughey, and B. Chesebro (2001)
J. Virol. 75, 10106-10112
   Abstract »    Full Text »    PDF »
Induction of Distinct [URE3] Yeast Prion Strains.
M. Schlumpberger, S. B. Prusiner, and I. Herskowitz (2001)
Mol. Cell. Biol. 21, 7035-7046
   Abstract »    Full Text »    PDF »
Acridine and phenothiazine derivatives as pharmacotherapeutics for prion disease.
C. Korth, B. C. H. May, F. E. Cohen, and S. B. Prusiner (2001)
PNAS 98, 9836-9841
   Abstract »    Full Text »    PDF »
Neurodegenerative Diseases and Prions.
S. B. Prusiner (2001)
N. Engl. J. Med. 344, 1516-1526
   Full Text »    PDF »
Branched Polyamines Cure Prion-Infected Neuroblastoma Cells.
S. Supattapone, H. Wille, L. Uyechi, J. Safar, P. Tremblay, F. C. Szoka, F. E. Cohen, S. B. Prusiner, and M. R. Scott (2001)
J. Virol. 75, 3453-3461
   Abstract »    Full Text »
Prion protein: Evolution caught en route.
P. Tompa, G. E. Tusnády, M. Cserz, and I. Simon (2001)
PNAS
   Abstract »    Full Text »
Molecular Analysis of the Abnormal Prion Protein during Coinfection of Mice by Bovine Spongiform Encephalopathy and a Scrapie Agent.
T. G. M. Baron and A.-G. Biacabe (2001)
J. Virol. 75, 107-114
   Abstract »    Full Text »
Affinity-Tagged Miniprion Derivatives Spontaneously Adopt Protease-Resistant Conformations.
S. Supattapone, H.-O. B. Nguyen, T. Muramoto, F. E. Cohen, S. J. DeArmond, S. B. Prusiner, and M. Scott (2000)
J. Virol. 74, 11928-11934
   Abstract »    Full Text »
Expression of unglycosylated mutated prion protein facilitates PrPSc formation in neuroblastoma cells infected with different prion strains.
C. Korth, K. Kaneko, and S. B. Prusiner (2000)
J. Gen. Virol. 81, 2555-2563
   Abstract »    Full Text »
Strain-specific propagation of PrPSc properties into baculovirus-expressed hamster PrPC.
V. Iniguez, D. McKenzie, J. Mirwald, and J. Aiken (2000)
J. Gen. Virol. 81, 2565-2571
   Abstract »    Full Text »
Genetic influence on the structural variations of the abnormal prion protein.
P. Parchi, W. Zou, W. Wang, P. Brown, S. Capellari, B. Ghetti, N. Kopp, W. J. Schulz-Schaeffer, H. A. Kretzschmar, M. W. Head, et al. (2000)
PNAS 97, 10168-10172
   Abstract »    Full Text »    PDF »
Species-barrier-independent prion replication in apparently resistant species.
A. F. Hill, S. Joiner, J. Linehan, M. Desbruslais, P. L. Lantos, and J. Collinge (2000)
PNAS 97, 10248-10253
   Abstract »    Full Text »    PDF »
Creutzfeldt-Jakob disease with a novel four extra-repeat insertional mutation in the PrP gene.
G. Rossi, G. Giaccone, L. Giampaolo, S. Iussich, G. Puoti, M. Frigo, G. Cavaletti, L. Frattola, O. Bugiani, and F. Tagliavini (2000)
Neurology 55, 405-410
   Abstract »    Full Text »    PDF »
Copper(II)-induced Conformational Changes and Protease Resistance in Recombinant and Cellular PrP. EFFECT OF PROTEIN AGE AND DEAMIDATION.
K. Qin, D.-S. Yang, Y. Yang, M. A. Chishti, L.-J. Meng, H. A. Kretzschmar, C. M. Yip, P. E. Fraser, and D. Westaway (2000)
J. Biol. Chem. 275, 19121-19131
   Abstract »    Full Text »    PDF »
Dominant-Negative Inhibition of Prion Formation Diminished by Deletion Mutagenesis of the Prion Protein.
L. Zulianello, K. Kaneko, M. Scott, S. Erpel, D. Han, F. E. Cohen, and S. B. Prusiner (2000)
J. Virol. 74, 4351-4360
   Abstract »    Full Text »
From the Cover: Compelling transgenetic evidence for transmission of bovine spongiform encephalopathy prions to humans.
M. R. Scott, R. Will, J. Ironside, H.-O. B. Nguyen, P. Tremblay, S. J. DeArmond, and S. B. Prusiner (1999)
PNAS 96, 15137-15142
   Abstract »    Full Text »    PDF »
Glycosylation differences between the normal and pathogenic prion protein isoforms.
P. M. Rudd, T. Endo, C. Colominas, D. Groth, S. F. Wheeler, D. J. Harvey, M. R. Wormald, H. Serban, S. B. Prusiner, A. Kobata, et al. (1999)
PNAS 96, 13044-13049
   Abstract »    Full Text »    PDF »
Type 1 protease resistant prion protein and valine homozygosity at codon 129 of PRNP identify a subtype of sporadic Creutzfeldt-Jakob disease.
B. B Worrall, S. T Herman, S. Capellari, T. Lynch, S. Chin, P. Gambetti, and P. Parchi (1999)
J. Neurol. Neurosurg. Psychiatry 67, 671-674
   Abstract »    Full Text »
Protease-resistant and Detergent-insoluble Prion Protein Is Not Necessarily Associated with Prion Infectivity.
G. M. Shaked, G. Fridlander, Z. Meiner, A. Taraboulos, and R. Gabizon (1999)
J. Biol. Chem. 274, 17981-17986
   Abstract »    Full Text »    PDF »
Prion Protein Conformation in a Patient with Sporadic Fatal Insomnia.
J. A. Mastrianni, R. Nixon, R. Layzer, G. C. Telling, D. Han, S. J. DeArmond, and S. B. Prusiner (1999)
N. Engl. J. Med. 340, 1630-1638
   Full Text »    PDF »
Molecular Genetics of Transmissible Spongiform Encephalopathies.
C. Weissmann (1999)
J. Biol. Chem. 274, 3-6
   Full Text »    PDF »
Protease-resistant prion protein produced in vitro lacks detectable infectivity.
A. Hill, M Antoniou, and J Collinge (1999)
J. Gen. Virol. 80, 11-14
   Abstract »
Strain-dependent Differences in beta -Sheet Conformations of Abnormal Prion Protein.
B. Caughey, G. J. Raymond, and R. A. Bessen (1998)
J. Biol. Chem. 273, 32230-32235
   Abstract »    Full Text »    PDF »
Prions.
S. B. Prusiner (1998)
PNAS 95, 13363-13383
   Abstract »    Full Text »    PDF »
Prions.
D. Westaway, G. Telling, and S. Priola (1998)
PNAS 95, 11030-11031
   Full Text »    PDF »
A Transmembrane Form of the Prion Protein in Neurodegenerative Disease.
R. S. Hegde, J. A. Mastrianni, M. R. Scott, K. A. DeFea, P. Tremblay, M. Torchia, S. J. DeArmond, S. B. Prusiner, and V. R. Lingappa (1998)
Science 279, 827-834
   Abstract »    Full Text »
Identification of a prion protein epitope modulating transmission of bovine spongiform encephalopathy prions to transgenic mice.
M. R. Scott, J. Safar, G. Telling, O. Nguyen, D. Groth, M. Torchia, R. Koehler, P. Tremblay, D. Walther, F. E. Cohen, et al. (1997)
PNAS 94, 14279-14284
   Abstract »    Full Text »    PDF »
Prion Diseases and the BSE Crisis.
S. B. Prusiner (1997)
Science 278, 245-251
   Abstract »    Full Text »
Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation.
K. Kaneko, L. Zulianello, M. Scott, C. M. Cooper, A. C. Wallace, T. L. James, F. E. Cohen, and S. B. Prusiner (1997)
PNAS 94, 10069-10074
   Abstract »    Full Text »    PDF »
Solution structure of a 142-residue recombinant prion protein corresponding to the infectious fragment of the scrapie isoform.
T. L. James, H. Liu, N. B. Ulyanov, S. Farr-Jones, H. Zhang, D. G. Donne, K. Kaneko, D. Groth, I. Mehlhorn, S. B. Prusiner, et al. (1997)
PNAS 94, 10086-10091
   Abstract »    Full Text »    PDF »
Evolution of a Strain of CJD That Induces BSE-Like Plaques.
L. Manuelidis, W. Fritch, and Y. Xi (1997)
Science 277, 94-98
   Abstract »    Full Text »
Scrapie susceptibility-linked polymorphisms modulate the in vitro conversion of sheep prion protein to protease-resistant forms.
A. Bossers, P. B. G. M. Belt, G. J. Raymond, B. Caughey, R. de Vries, and M. A. Smits (1997)
PNAS 94, 4931-4936
   Abstract »    Full Text »    PDF »
Identification of Intermediate Steps in the Conversion of a Mutant Prion Protein to a Scrapie-like Form in Cultured Cells.
N. Daude, S. Lehmann, and D. A. Harris (1997)
J. Biol. Chem. 272, 11604-11612
   Abstract »    Full Text »    PDF »
pH-dependent Prion Protein Conformation in Classical Creutzfeldt-Jakob Disease.
G. Zanusso, A. Farinazzo, M. Fiorini, M. Gelati, A. Castagna, P. G. Righetti, N. Rizzuto, and S. Monaco (2001)
J. Biol. Chem. 276, 40377-40380
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

ADVERTISEMENT

Featured Jobs

Science. ISSN 0036-8075 (print), 1095-9203 (online)