Cell, Vol.
93, 337–348, May 1, 1998, Copyright 1998 by Cell Press
Prion Protein Biology
Stanley B. Prusiner,*† Michael R. Scott,*
Stephen J. DeArmond,‡* and Fred E. Cohen†§k
* Department of Neurology
† Department of Biochemistry and Biophysics
‡ Department of Pathology
§ Department of Molecular and Cellular Pharmacology
k Department of Medicine
University of California
San Francisco, California 94143
Introduction
It is interesting to contemplate how the course of scien-
tific investigation might have proceeded had studies on
the transmissibility of inherited prion diseases not been
performed until after the molecular genetic lesion had
been identified (Meggendorfer, 1930; Roos et al., 1973;
Hsiao et al., 1989). Had the prion protein (PrP) gene been
identified in families with prion disease by positional
cloning or through the purification and sequencing of
PrP in amyloid plaques before brain extracts were
shown to be transmissible, the prion concept might have
been more readily accepted (Prusiner, 1998).
But that is not the course of events that led to our
current understanding of prions. Creutzfeldt-Jakob dis-
ease (CJD) remained a curious, rare neurodegenerative
disease of unknown etiology for more than three score
years. Only the transmission of CJD to apes by inocula-
tion of brain extracts from patients who had died of CJD
initiated a path of scientific investigation that was to
demystify that fascinating area of biomedicine (Gibbs
et al., 1968). Not unexpectedly, once CJD was shown
to be an infectious disease, relatively little attention was
paid to the familial form of the disease since most cases
are sporadic and familial clusters account for a minority.
In this review, we focus on the prion particles that
cause scrapie, bovine spongiform encephalopathy (BSE),
and CJD. Some of the fascinating studies that led to
our knowledge of prions are not discussed here but
have recently been reviewed elsewhere (Prusiner, 1998).
Prion diseases may present as genetic, infectious, or
sporadic disorders, all of which involve modification of
the prion protein (PrP), a constituent of normal mamma-
lian cells. CJD generally presents as progressive demen-
tia while scrapie of sheep and BSE are generally mani-
fest as ataxic illnesses (Table 1) (Wells et al., 1987). In
CJD, scrapie, and BSE, as well as all of the other disor-
ders frequently referred to as prion diseases (Table 1),
spongiform degeneration and astrocytic gliosis are
found upon microscopic examination of the CNS. The
degree of spongiform degeneration is quite variable
while the extent of reactive gliosis correlates with the
degree of neuron loss (Masters and Richardson, 1978).
The Prion Particle
Perhaps, the best current working definition of a prion
is a proteinaceous infectious particle that lacks nucleic
acid (Prusiner, 1997). A wealth of data supports the
contention that scrapie prions are devoid of nucleic acid
Review
and seem to be composed exclusively of a modified
isoform of PrP, designated PrPSc. The normal cellular
PrP, denoted PrPC, is converted into PrPSc through a
process whereby a portion of its a-helical and coil struc-
ture is refolded into b sheet (Pan et al., 1993). This struc-
tural transition is accompanied by profound changes in
the physicochemical properties of the PrP. While PrPC
is soluble in nondenaturing detergents, PrPSc is not. PrPC
is readily digested by proteases, whereas PrPSc is par-
tially resistant (Oesch et al., 1985). Because prions ap-
pear to be composed entirely of a protein that adopts
an abnormal conformation, it is not unreasonable to
think of prions as infectious proteins (Pan et al., 1993;
Telling et al., 1996). But we hasten to add that we still
cannot eliminate the possibility of a small ligand bound
to PrPSc as an essential component of the infectious
prion particle.
In a more broad view, prions are elements that impart
and propagate variability through multiple conformers
of a normal cellular protein. The species of a particular
prion is encoded by the sequence of the chromosomal
PrP gene of the mammal in which it last replicated. In
contrast to pathogens with a nucleic acid genome that
encode strain-specific properties in genes, prions seem
to encipher these properties in the tertiary structure
of PrPSc (Bessen and Marsh, 1994; Telling et al., 1996;
Prusiner, 1997).
The discovery that mutations of the PrP gene caused
dominantly inherited prion diseases in humans linked
the genetic and infectious forms of prion diseases and
presented another hurdle for investigators who contin-
ued to argue that prion diseases are caused by viruses.
More than 20 mutations of the PrP gene are now known
to cause the inherited human prion diseases, and signifi-
cant genetic linkage has been established for five of
these mutations (Hsiao et al., 1989) (for review, see Prus-
iner, 1997). The prion concept readily explains how a
disease can be manifest as a heritable as well as an
infectious illness. Moreover, the hallmark common to
all of the prion diseases whether sporadic, dominantly
inherited, or acquired by infection is that they involve
the aberrant metabolism of the prion protein (Prusiner,
1991).
Although PrPSc is the only known component of the
infectious prion particles, these unique pathogens share
several phenotypic traits with other infectious entities
such as viruses. Because some features of the diseases
caused by prions and viruses are similar, some scien-
tists have difficulty accepting the existence of prions
despite a wealth of scientific data supporting this con-
cept (Chesebro and Caughey, 1993; Manuelidis and
Fritch, 1996; Lasmézas et al., 1997; Chesebro, 1998).
Molecular Genetics of Prion Diseases
Once a PrP cDNA probe became available, molecular
genetic studies were undertaken to determine whether
the PrP gene controls scrapie incubation times in mice.
Independent of the enriching of brain fractions for
scrapie infectivity that led to the discovery of PrPSc, the
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Table 1. The Prion Diseases

Disease Host Mechanism of Pathogenesis

Kuru Fore people Infection through ritualistic cannibalism
iCJD Humans Infection from prion-contaminated HGH, dura mater
grafts, etc.
vCJD Humans Infection from bovine prions?
fCJD Humans Germline mutations in PrP gene
GSS Humans Germline mutations in PrP gene
FFI Humans Germline mutation in PrP gene (D178N, M129)
sCJD Humans Somatic mutation or spontaneous conversion of PrP C
into PrP Sc?
FSI Humans Somatic mutation or spontaneous conversion of PrP C
into PrP Sc?
Scrapie Sheep Infection in genetically susceptible sheep
BSE Cattle Infection with prion-contaminated MBM
TME Mink Infection with prions from sheep or cattle
CWD Mule deer, elk Unknown
FSE Cats Infection with prion-contaminated beef
Exotic ungulate encephalopathy Greater kudu, nyala, oryx Infection with prion-contaminated MBM

Abbreviations: BSE, bovine spongiform encephalopathy; CJD, Creutzfeldt-Jakob disease; sCJD, sporadic CJD; fCJD, familial CJD; iCJD,
iatrogenic CJD; vCJD, (new) variant CJD; CWD, chronic wasting disease; FFI, fatal familial insomnia; FSE, feline spongiform encephalopathy;
FSI, fatal sporadic insomnia; GSS, Gerstmann-Sträussler-Scheinker disease; HGH, human growth hormone; MBM, meat and bone meal; TME,
transmissible mink encephalopathy.

PrP gene was shown to be genetically linked to a locus
controlling the incubation time (Carlson et al., 1986).
Subsequently, mutation of the PrP gene was shown to
be genetically linked to the development of familial prion
disease (Hsiao et al., 1989). At the same time, expression
of a Syrian hamster (SHa) PrP transgene in mice was
shown to render the animals highly susceptible to SHa
prions, which demonstrated that expression of a foreign
PrP gene could abrogate the species barrier (Scott et
al., 1989). Later, PrP-deficient (Prnp 0/0) mice were found
to be resistant to prion infection and failed to replicate
prions, as expected (Büeler et al., 1993; Prusiner et al.,
1993). The results of these studies indicated PrP must
play a central role in the transmission and pathogenesis
of prion disease, but equally important, they established
that the abnormal isoform is an essential component of
the prion particle (Prusiner, 1991).
PrP Gene Dosage Controls Length
of Incubation Time
Scrapie incubation times in mice were used to distin-
guish prion strains and to identify a gene controlling its
length (Dickinson et al., 1968; Scott et al., 1997). This
gene was initially called Sinc based on genetic crosses
between C57Bl and VM mice, which exhibited short and
long incubation times, respectively (Dickinson et al.,
1968). Because the distribution of VM mice was re-
stricted, we searched for another mouse with long incu-
bation times. I/Ln mice proved to be a suitable substitute
for VM mice; eventually, I/Ln and VM mice were found
to be derived from a common ancestor. Subsequently,
the PrP gene was shown to control the length of the
scrapie incubation time in mice (Carlson et al., 1994;
Moore et al., 1998).
Overexpression of wtPrP Transgenes
Mice were constructed expressing different levels of the
wild-type (wt) SHaPrP transgene (Scott et al., 1989).
Inoculation of these Tg(SHaPrP) mice with SHa prions
demonstrated abrogation of the species barrier resulting
in abbreviated incubation times due to a nonstochastic
process (Prusiner et al., 1990). The length of the incuba-
tion time after inoculation with SHa prions was inversely
proportional to the level of SHaPrPC in the brains of
Tg(SHaPrP) mice (Prusiner et al., 1990). Bioassays of
brain extracts from clinically ill Tg(SHaPrP) mice inocu-
lated with mouse (Mo) prions revealed that only Mo
prions but no SHa prions were produced. Conversely,
inoculation of Tg(SHaPrP) mice with SHa prions led to
the synthesis of only SHa prions. Thus, the rate of PrPSc
synthesis appears to be a function of the level of PrPC
expression in Tg mice; however, the level to which PrPSc
accumulates appears to be independent of PrPC con-
centration (Prusiner et al., 1990).
PrP-Deficient Mice
The development and lifespan of two lines of Prnp0/0
mice were indistinguishable from controls (Büeler et al.,
1992; Manson et al., 1994) while another line exhibited
ataxia and Purkinje cell degeneration at z70 weeks of
age (Sakaguchi et al., 1996). In the former two lines with
normal development, altered sleep–wake cycles (Tobler
et al., 1996) and synaptic behavior in brain slices have
been reported (Collinge et al., 1994), but the synaptic
changes could not be confirmed by others (Lledo et al.,
1996).
Prnp0/0 mice are resistant to prions (Büeler et al., 1993;
Prusiner et al., 1993). Prnp0/0 mice were sacrificed 5, 60,
120, and 315 days after inoculation with RML prions and
brain extracts bioassayed in CD-1 Swiss mice. Except
for residual infectivity from the inoculum detected at 5
days after inoculation, no infectivity was detected in the
brains of Prnp0/0 mice (Prusiner et al., 1993). One group
of investigators found that Prnp0/0 mice inoculated with
RML prions and sacrificed 20 weeks later had 103.6 ID50
units/ml of homogenate by bioassay (Büeler et al., 1993).
Others have used this report to argue that prion infectiv-
ity replicates in the absence of PrP (Chesebro and
Caughey, 1993; Lasmézas et al., 1997). Neither we nor
the authors of the initial report could confirm the finding
of prion replication in Prnp0/0 mice (Prusiner et al., 1993;
Sailer et al., 1994).
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339

Prion Protein Structure

Once cDNA probes for PrP became available, the PrP
gene was found to be constitutively expressed in adult,
uninfected brain (Chesebro et al., 1985; Oesch et al.,
1985). This finding eliminated the possibility that PrPSc
stimulated production of more of itself by initiating tran-
scription of the PrP gene as proposed nearly two de-
cades earlier (Griffith, 1967). Determination of the struc-
ture of the PrP gene eliminated a second possible
mechanism that might explain the appearance of PrPSc
in brains already synthesizing PrPC. Since the entire pro-
tein coding region was contained within a single exon,
there was no possibility that the two PrP isoforms were
the products of alternatively spliced mRNAs (Basler et
al., 1986). Next, a posttranslational chemical modifica-
tion that distinguishes PrPSc from PrPC was considered
but none was found in an exhaustive study (Stahl et al.,
1993), and we considered it likely that PrPC and PrPSc
differed only in their conformations, a hypothesis also
proposed earlier (Griffith, 1967).
When the secondary structures of the PrP isoforms
were compared by optical spectroscopy, they were
found to be markedly different (Pan et al., 1993). Fourier
transform infrared (FTIR) and circular dichroism (CD)
spectroscopy studies showed that PrPC contains about
40% a helix and little b sheet while PrPSc is composed
of about 30% a helix and 45% b sheet (Pan et al., 1993).
That the two PrP isoforms have the same amino acid
sequence runs counter to the widely accepted view that
Figure 1. Species Variations and Mutations of the Prion Protein
the amino acid sequence specifies only one biologically Gene
active conformation of a protein (Anfinsen, 1973). (A) Species variations. The x-axis represents the human PrP se-
Prior to comparative studies on the structures of PrPC quence, with the five octarepeats and H1–H4 regions of putative
and PrPSc, metabolic labeling studies showed that the secondary structure shown as well as the three a helices A, B, and
acquisition of PrPSc protease resistance is a posttransla- C and the two b strands S1 and S2. Vertical bars above the axis
tional process (Borchelt et al., 1990). In a search for indicate the number of species that differ from the human sequence
at each position. Below the axis, the length of the bars indicates the
chemical differences that would distinguish PrPSc from
number of alternative amino acids at each position in the alignment.
PrPC, we identified ethanolamine in hydrolysates of PrP (B) Mutations causing inherited human prion disease and polymor-
27–30, which signaled the possibility that PrP might con- phisms in human, mouse, and sheep. Above the line of the human
tain a glycosylphosphatidyl inositol (GPI) anchor (Stahl sequence are mutations that cause prion disease. Below the lines
et al., 1987). Both PrP isoforms were found to carry GPI are polymorphisms, some but not all of which are known to influence
anchors and PrPC was found on the surface of cells the onset as well as the phenotype of disease. Data were compiled
by Paul Bamborough and Fred E. Cohen. Reprinted with permission
where it could be released by cleavage of the anchor.
from Science 278, pp. 245–251 (copyright 1997 American Associa-
Subsequent studies showed that PrPSc formation occurs tion for the Advancement of Science).
after PrPC reaches the cell surface (Caughey and Ray-
mond, 1991) and is localized to caveolae-like domains
(Gorodinsky and Harris, 1995; Taraboulos et al., 1995). these recombinant PrPs correspond to the secreted
Computational Models and Optical Spectroscopy form of PrP that was also identified in the cell-free trans-
Modeling studies and subsequent nuclear magnetic res- lation studies. This contention is supported by studies
onance (NMR) investigations of a synthetic PrP peptide with recombinant antibody fragments (Fabs) showing
containing residues 90–145 suggested that PrPC might that GPI-anchored PrPC on the surface of cells exhibits
contain an a helix within this region (Figure 1) (Huang an immunoreactivity similar to that of recombinant PrP
et al., 1994). This peptide contains the residues, 113– prepared with an a-helical conformation (Peretz et al.,
128, that are most highly conserved among all species 1997).
studied (Figure 1A) and that correspond to a transmem- Models of PrPSc suggest that formation of the disease-
brane region of PrP which was delineated in cell-free causing isoform involves refolding of a region corre-
translation studies. A transmembrane form of PrP was sponding roughly to residues 108–144 into b sheets (Hu-
found in brains of patients with Gerstmann-Sträussler- ang et al., 1996); the single disulfide bond joining the
Scheinker disease (GSS) caused by the A117V mutation COOH-terminal helices would remain intact since the
and in Tg mice overexpressing either mutant or wtPrP disulfide is required for PrPSc formation (Figure 2D) (Mur-
(Hegde et al., 1998). That no evidence for an a helix in amoto et al., 1996). Deletion of each of several regions
this region has been found in NMR studies of recombi- of putative secondary structure in PrP, except for the
nant PrP in an aqueous environment (Riek et al., 1996; NH 2-terminal 66 amino acids (residues 23–88) and a 36–
Donne et al., 1997; James et al., 1997) suggests that amino acid stretch (Mo residues 141–176), prevented
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Figure 2. Structures of Prion Proteins

(A) NMR structure of Syrian hamster (SHa) recombinant (r) PrP(90–231). Presumably, the structure of the a-helical form of rPrP(90–231)
resembles that of PrP C. rPrP(90–231) is viewed from the interface where PrPSc is thought to bind to PrPC. Color scheme: a helices A (residues
144–157), B (172–193), and C (200–227) in pink; disulfide between Cys-179 and Cys-214 in yellow; conserved hydrophobic region composed
of residues 113–126 in red; loops in gray; residues 129–134 in green encompassing strand S1 and residues 159–165 in blue encompassing
strand S2; the arrows span residues 129–131 and 161–163, as these show a closer resemblance to b sheet (James et al., 1997).
(B) NMR structure of rPrP(90–231) is viewed from the interface where protein X is thought to bind to PrPC. Protein X appears to bind to the
side chains of residues that form a discontinuous epitope: some amino acids are in the loop composed of residues 165–171 and at the end
of helix B (Gln-168 and Gln-172 with a low density van der Waals rendering) while others are on the surface of helix C (Thr-215 and Gln-219
with a high density van der Waals rendering) (Kaneko et al., 1997b).
(C) Schematic diagram showing the flexibility of the polypeptide chain for PrP(29–231) (Donne et al., 1997). The structure of the portion of the
protein representing residues 90–231 was taken from the coordinates of PrP(90–231) (James et al., 1997). The remainder of the sequence
was hand-built for illustration purposes only. The color scale corresponds to the heteronuclear { 1H}-15N NOE data: red for the lowest (most
negative) values, where the polypeptide is most flexible, to blue for the highest (most positive) values in the most structured and rigid regions
of the protein. Reprinted with permission from Proc. Natl. Acad. Sci. USA 94, pp. 13452–13457 (copyright 1997 National Academy of Sciences).
(D) Plausible model for the tertiary structure of human PrPSc (Huang et al., 1996). Color scheme: S1 b strands are 108–113 and 116–122 in
red; S2 b strands are 128–135 and 138–144 in green; a helices H3 (residues 178–191) and H4 (residues 202–218) in gray; loop (residues
142–177) in yellow. Four residues implicated in the species barrier are shown in ball-and-stick form (Asn-108, Met-112, Met-129, Ala-133).
Reprinted with permission from Science 278, pp. 245–251 (copyright 1997 American Association for the Advancement of Science).

formation of PrPSc as measured in scrapie-infected cul-
tured neuroblastoma cells (Muramoto et al., 1996). With
a-PrP Fabs selected from phage display libraries and
two monoclonal antibodies (MAbs) derived from hybrid-
omas, a major conformational change that occurs during
conversion of PrPC into PrPSc has been localized to resi-
dues 90–112 (Peretz et al., 1997). Studies with an a-PrP
IgM MAb, which was reported to immunoprecipitate
PrP Sc selectively (Korth et al., 1997), support this conclu-
sion. While these results indicate that PrPSc formation
involves a conformational change at the NH 2 terminus,
mutations causing inherited prion diseases have been
found throughout the protein (Figure 1B). Interestingly,
all of the known point mutations in PrP with biological
significance occur either within or adjacent to regions
of putative secondary structure in PrP and, as such,
appear to destabilize the structure of PrPC (Huang et al.,
1994; Riek et al., 1996).
NMR Structure of Recombinant PrP
The NMR structure of recombinant SHaPrP(90–231) was
determined after the protein was purified and refolded
(Figure 2A). Residues 90–112 are not shown since
marked conformational heterogeneity was found in this
region while residues 113–126 constitute the conserved
hydrophobic region that also displays some structural
plasticity (James et al., 1997). Although some features
of the structure of rPrP(90–231) are similar to those re-
ported earlier for the smaller recombinant MoPrP(121–
231) fragment (Riek et al., 1996), substantial differences
were found. For example, the loop at the NH2 terminus
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341
into a nascent molecule of PrP Sc, protein X is released and a dimer of PrPSc remains. The inactivation target size of an infectious prion suggests
that it is composed of a dimer of PrP Sc (Bellinger-Kawahara et al., 1988). In the model depicted here, a fraction of infectious PrP Sc dimers
dissociate into uninfectious monomers as the replication cycle proceeds while most of the dimers accumulate in accord with the increase in
prion titer that occurs during the incubation period. The precise stoichiometry of the replication process remains uncertain.
of helix B is defined in rPrP(90–231) but is disordered
in MoPrP(121–231); in addition, helix C is composed of
residues 200–227 in rPrP(90–231) but extends only from
200–217 in MoPrP(121–231). The loop and the COOH-
terminal portion of helix C are particularly important as
described below (Figure 2B). Whether the differences
between the two recombinant PrP fragments are due
to (i) their different lengths, (ii) species-specific differ-
ences in sequences, or (iii) the conditions used for solv-
ing the structures remains to be determined.
Recent NMR studies of full-length MoPrP(23–231) and
SHaPrP(29–231) have shown that the NH2 termini are
highly flexible and lack identifiable secondary structure
under the experimental conditions employed (Figure 2C)
(Donne et al., 1997). Studies of SHaPrP(29–231) indicate
transient interactions between the COOH-terminal end
of helix B and the highly flexible, NH2-terminal ran-
dom-coil containing the octareapeats (residues 29–125)
(Donne et al., 1997).
Prion Replication
In an uninfected cell, PrP C with the wild-type sequence
exists in equilibrium in its monomeric a-helical, prote-
ase-sensitive state or bound to protein X (Figure 3). We
denote the conformation of PrPC that is bound to pro-
tein X as PrP * (Cohen et al., 1994); this conformation is
likely to be different from that determined under aque-
ous conditions for monomeric recombinant PrP. The
PrP* /protein X complex will bind PrPSc, thereby creating
a replication-competent assembly. Order of addition ex-
periments demonstrate that for PrPC, protein X binding
precedes productive PrPSc interactions (Kaneko et al.,
1997b). A conformational change takes place wherein
PrP, in a shape competent for binding to protein X and
PrPSc , represents the initial phase in the formation of
infectious PrPSc.
Several lines of evidence argue that the smallest infec-
tious prion particle is an oligomer of PrPSc , perhaps as
small as a dimer (Bellinger-Kawahara et al., 1988). Upon
purification, PrPSc tends to aggregate into insoluble
multimers that can be dispersed into liposomes (Gabi-
zon et al., 1988). Insolubility does not seem to be a
prerequisite for PrPSc formation or prion infectivity,
as suggested by some investigators (Gajdusek, 1988;
Caughey et al., 1995); a protease-resistant PrPSc that is
soluble in 1% Sarkosyl was generated in ScN2a cells
by expression of a PrP deletion mutant consisting of
106 amino acid residues (Muramoto et al., 1996).
In attempts to form PrPSc in vitro, PrPC has been ex-
posed to 3 M guanidinium HCl and then diluted 10-fold
prior to binding to PrPSc (Kocisko et al., 1994; Kaneko
et al., 1997a). Based on these results, we presume that
Figure 3. Schematic Diagram Showing Tem-
plate-Assisted PrPSc Formation
In the initial step, PrPC binds to protein X to
form the PrP */protein X complex. Next, PrP Sc
binds to PrP* that has already formed a com-
plex with protein X. When PrP* is transformed
exposure of PrPC to GdnHCl converts it into a PrP*-like
molecule. Whether this PrP* -like protein is converted
into PrPSc is unclear. Although the PrP*-like protein
bound to PrPSc is protease-resistant and insoluble, this
protease-resistant PrP has not been reisolated in order
to assess whether or not it was converted into PrPSc. It
is noteworthy that recombinant PrP can be refolded into
either a-helical or b-sheet forms but none have been
found to possess prion infectivity as judged by bioassay.
Inherited and Sporadic Prion Diseases
For inherited and sporadic prion diseases, the major
question is how the first PrPSc molecules are formed.
Once these are formed, replication presumably follows
the mechanism outlined for infectious disease. Several
lines of evidence suggest that PrPSc is more stable than
PrPC and that a kinetic barrier precludes the formation
of PrPSc under normal conditions. In the case of the
initiation of inherited prion diseases, the barrier to PrPSc
formation must be lower for the mutant (DPrPC) than the
wild-type and thus DPrP* can spontaneously rearrange
to form DPrPSc. While the known mutations would appear
to be destabilizing to the structure of PrPC, we lack
useful information about the structure of the transition
state for either the mutant or wild-type sequences. Stud-
ies of PrP in the brains of patients who were heterozy-
gous for the E200K mutation revealed DPrPSc(E200K)
molecules that were both detergent-insoluble and re-
sistant to limited proteolysis while most wtPrP was
detergent-insoluble but protease-sensitive (Gabizon
et al., 1996). These results suggest that in familial (f)
CJD(E200K), insoluble wtPrP might represent a form of
PrP* (Gabizon et al., 1996). In studies with CHO cells,
expression of DPrP(E200K) was found to be accompa-
nied with the posttranslational acquisition of resistance
to limited proteolysis (Lehmann and Harris, 1996), but
whether such cell lines expressing DPrP(E200K) pro-
duce infectious prions is unknown. It is noteworthy that
levels of proteinase K used in the studies where
DPrP(E200K) was expressed in CHO cells were lower
by a factor of 10–100 compared to digestions of PrPSc
derived from brain or ScN2a cells. Whether these alter-
ations in the properties of DPrP(E200K) in CHO cells
provide evidence for DPrP* or such changes lie outside
the pathway of DPrPSc(E200K) formation remains to be
determined.
Initiation of sporadic disease may follow from a so-
matic mutation and thus follow a path similar to that for
germline mutations in inherited disease. In this situation,
the mutant PrPSc must be capable of co-opting wtPrPC,
a process known to be possible for some mutations
(e.g., E200K, D178N) but less likely for others (e.g.,
P102L) (Telling et al., 1995, 1996). Alternatively, the acti-
vation barrier separating wtPrP C from PrPSc could be
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crossed on rare occasions when viewed in the context
of a population. Most individuals would be spared while
presentations in the elderly with an incidence of z1 per
million would be seen.
Mechanism of Prion Propagation?
From the foregoing formalism, we can ask “What is the
rate-limiting step in prion formation?” First, we must
consider the impact of the concentration of PrPSc in the
inoculum, which is inversely proportional to the length
of the incubation time. Second, we must consider the
sequence of PrPSc that forms an interface with PrPC.
When the sequences of the two isoforms are identical,
the shortest incubation times are observed. Third, we
must consider the strain-specific conformation of PrPSc.
Some prion strains exhibit longer incubation times than
others; the mechanism underlying this phenomenon is
not understood. From these considerations, there exists
a set of conditions under which initial PrPSc concentra-
tions can be rate-limiting. These effects presumably re-
late to the stability of the PrPSc, its targeting to the correct
cells and subcellular compartments, and its ability to be
cleared. Once infection in a cell is initiated and endoge-
nous PrPSc production is operative, then the following
discussion of PrPSc formation seems most applicable.
If the assembly of PrPSc into a specific dimeric or
multimeric arrangement were difficult, then a nucle-
ation–polymerization (NP) formalism would be relevant.
In NP processes, nucleation is the rate-limiting step and
elongation or polymerization is facile. These conditions
are frequently observed in peptide models of aggrega-
tion phenomena (Caughey et al., 1995); however, studies
with Tg mice expressing foreign PrP genes suggest that
a different process is occurring. From investigations
with mice expressing both the SHaPrP transgene and
the endogenous MoPrP gene, it is clear that PrPSc pro-
vides a template for directing prion replication where
we define a template as a catalyst that leaves its im-
print on the product of the reaction (Prusiner et al.,
1990). Inoculation of these mice with SHaPrPSc leads to
the production of nascent SHaPrPSc and not MoPrPSc.
Conversely, inoculation of the Tg(SHaPrP) mice with
MoPrPSc results in MoPrPSc formation and not SHaPrPSc.
Even stronger evidence for templating has emerged from
studies of prion strains passaged in Tg(MHu2M)Prnp0/0
mice expressing a chimeric Hu/MoPrP gene as de-
scribed in more detail below (Telling et al., 1996; Prusi-
ner, 1997). Even though the conformational templates
were initially generated with PrPSc molecules having dif-
ferent sequences in patients with inherited prion dis-
eases, these templates are sufficient to direct replication
of distinct PrPSc molecules when the amino acid se-
quences of the substrate PrPs are identical. If the forma-
tion of this template were rate-limiting, then an NP model
could apply. However, studies of PrPSc formation in
ScN2a cells point to a distinct rate-limiting step.
Cell biologic and transgenetic investigations argue for
the existence of a chaperone-like molecule, referred to
as protein X, that is required for PrPSc formation (Telling
et al., 1995). As described below, mutagenesis experi-
ments have created dominant negative forms of DPrPC
that inhibit the formation of wtPrPSc by binding protein X
(Kaneko et al., 1997b). This implies that the rate-limiting
step in vivo in prion replication under conditions where
Table 2. Evidence for Protein X from Transmission Studies of
Human Prionsa
Inoculum Host MoPrP Incubation Time
Gene [days 6 SEM] (n/no)
sCJD Tg(HuPrP) Prnp1/1 721 (1/10)
sCJD Tg(HuPrP)Prnp0/0 Prnp0/0 263 6 2 (6/6)
sCJD Tg(MHu2M) Prnp1/1 238 6 3 (8/8)
sCJD Tg(MHu2M)Prnp0/0 Prnp0/0 191 6 3 (10/10)
a
Data with inoculum RG from Telling et al., 1995.
PrPSc is sufficient must be the conversion of PrPC to PrP*
since a dominant negative derived from a single point
mutation could gate only a kinetically critical step in a
cellular process. In the template-directed model, the
conversion of PrPC to PrP* is a first order process. By
contrast, NP processes follow higher order kinetics
([monomer]m, where m is the number of monomers in
the nucleus). The experimental implications of these rate
relationships are apparent in transgenic studies; if first
order kinetics operate, halving the gene dose (hemizy-
gotes) should double the incubation time while doubling
the dose of a transgene array should halve the time to
disease. This quantitative behavior has been observed
in several studies in mice with altered levels of PrP
expression (Prusiner et al., 1990, 1993; Büeler et al.,
1994; Carlson et al., 1994). The existence of prion strains
that are conformational isoforms of PrPSc with distinct
structures, incubation times, and neurohistopathology
must also be considered in an analysis of the kinetics
of PrPSc accumulation. Since the rate-limiting step in
PrPSc formation cannot involve the unique template pro-
vided by a strain, differential rates of intercellular spread,
cellular uptake, and clearance seem most likely to ac-
count for the variation in incubation times. This is consis-
tent with the different patterns of protease sensitivity
and glycosylation for distinct prion strains (Bessen and
Marsh, 1994; Collinge et al., 1996; Telling et al., 1996;
Somerville et al., 1997).
However, we hasten to add that NP models can pro-
vide a useful description of other biologic phenomena.
Under conditions when the monomer is relatively rare
and/or the conformational change is facile (e.g., short
peptides), the NP model will dominate. However, when
the monomer is sufficiently abundant and/or the confor-
mational conversion is difficult to accomplish, the tem-
plate assistance formalism provides a more likely de-
scription of the process.
Evidence for Protein X
Protein X was postulated to explain the results on the
transmission of human (Hu) prions to Tg mice (Table 2)
(Telling et al., 1994, 1995). Mice expressing both Mo and
HuPrP were resistant to Hu prions while those express-
ing only HuPrP were susceptible. These results argue
that MoPrPC inhibited transmission of Hu prions, i.e.,
the formation of nascent HuPrP Sc. In contrast to the
foregoing studies, mice expressing both MoPrP and chi-
meric MHu2M PrP were susceptible to Hu prions and
mice expressing MHu2MPrP alone were only slightly
more susceptible. These findings contend that MoPrPC
has only a minimal effect on the formation of chimeric
MHu2MPrPSc.
Review
343
When the data on Hu prion transmission to Tg mice
were considered together, they suggested that MoPrPC
prevented the conversion of HuPrPC into PrPSc by bind-
ing to another Mo protein but had little effect on the
conversion of MHu2M into PrPSc. We interpreted these
results in terms of MoPrPC binding to this Mo protein
with a higher affinity than does HuPrP C. We postulated
that MoPrPC had little effect on the formation of PrPSc
from MHu2M (Table 2) because MoPrP and MHu2M
share the same amino acid sequence at the COOH termi-
nus. This also suggested that MoPrPC only weakly inhib-
ited transmission of SHa prions to Tg(SHaPrP) mice be-
cause SHaPrP is more closely related to MoPrP than is
HuPrP.
Using scrapie-infected mouse neuroblastoma cells
transfected with chimeric Hu/Mo PrP genes, we ex-
tended our studies of protein X. Substitution of a Hu
residue at position 214 or 218 prevented PrPSc formation
(Figure 2B) (Kaneko et al., 1997b). The side chains of
these residues protrude from the same surface of the
COOH-terminal a helix forming a discontinuous epitope
with residues 167 and 171 in an adjacent loop. Substitu-
tion of a basic residue at positions 167, 171, or 218
prevented PrPSc formation; these mutant PrPs appear
to act as “dominant negatives” by binding protein X
and rendering it unavailable for prion propagation. Our
findings seem to explain the protective effects of basic
polymorphic residues in PrP of humans and sheep
(Hunter et al., 1993; Westaway et al., 1994; Shibuya et
al., 1998).
Is Protein X a Molecular Chaperone?
Since PrP undergoes a profound structural transition
during prion propagation, it seems likely that other pro-
teins such as chaperones participate in this process.
Whether protein X functions as a classical molecular
chaperone or participates in PrP binding as part of its
normal function but can also facilitate pathogenic as-
pects of PrP biology is unknown. Interestingly, scrapie-
infected cells in culture display marked differences in
the induction of heat-shock proteins (Tatzelt et al., 1995),
and Hsp70 mRNA has been reported to increase in
scrapie of mice (Kenward et al., 1994). While attempts
to isolate specific proteins that bind to PrP have been
disappointing (Oesch et al., 1990), PrP has been shown
to interact with Bcl-2, Hsp60, and the laminin receptor
protein by two-hybrid analysis in yeast (Kurschner and
Morgan, 1996; Rieger et al., 1997). Although these stud-
ies are suggestive, no molecular chaperone involved in
prion formation in mammalian cells has been identified.
Strains of Prions
The existence of prion strains raises the question of how
heritable biological information can be enciphered in
any molecule other than nucleic acid (Dickinson et al.,
1968). Strains or varieties of prions have been defined
by incubation times and the distribution of neuronal vac-
uolation (Dickinson et al., 1968; Fraser and Dickinson,
1968). Subsequently, the patterns of PrPSc deposition
were found to correlate with vacuolation profiles, and
these patterns were also used to characterize strains of
prions (DeArmond et al., 1987, 1997; Bruce et al., 1989).
The typing of prion strains in C57Bl, VM, and
F1(C57Bl 3 VM) inbred mice began with isolates from
sheep with scrapie. The prototypic strains called Me7
and 22A gave incubation times of z150 and z400 days
in C57Bl mice, respectively (Dickinson et al., 1968). The
PrPs of C57Bl and I/Ln (and later VM) mice differ at two
residues and control incubation times (Carlson et al.,
1994; Moore et al., 1998).
Until recently, support for the hypothesis that the ter-
tiary structure of PrPSc enciphers strain-specific informa-
tion (Prusiner, 1991) was minimal except for the DY strain
isolated from mink with transmissible encephalopathy
(Bessen and Marsh, 1994). PrPSc in DY prions showed
diminished resistance to proteinase K digestion as well
as an anomalous site of cleavage. The DY strain pre-
sented a puzzling anomaly since other prion strains ex-
hibiting similar incubation times did not show this al-
tered susceptibility to proteinase K digestion of PrP Sc
(Scott et al., 1997). Also notable was the generation of
new strains during passage of prions through animals
with different PrP genes (Scott et al., 1997).
PrP Sc Conformation Enciphers Variation in Prions
Persuasive evidence that strain-specific information is
enciphered in the tertiary structure of PrPSc comes from
transmission of two different inherited human prion
diseases to mice expressing a chimeric MHu2M PrP
transgene (Telling et al., 1996). In fatal familial insomnia
(FFI), the protease-resistant fragment of PrPSc after
deglycosylation has an Mr of 19 kDa; whereas in
fCJD(E200K) and most sporadic prion diseases, it is 21
kDa (Monari et al., 1994). This difference in molecular
size was shown to be due to different sites of proteolytic
cleavage at the NH 2 termini of the two human PrPSc
molecules reflecting different tertiary structures (Monari
et al., 1994). These distinct conformations were not un-
expected since the amino acid sequences of the PrPs
differ.
Extracts from the brains of FFI patients transmitted
disease into mice expressing a chimeric MHu2M PrP
gene about 200 days after inoculation and induced for-
mation of the 19 kDa PrPSc ; whereas fCJD(E200K) and
sCJD produced the 21 kDa PrPSc in mice expressing
the same transgene (Telling et al., 1996). On second
passage, Tg(MHu2M) mice inoculated with FFI prions
showed an incubation time of z130 days and a 19 kDa
PrP Sc while those inoculated with fCJD(E200K) prions
exhibited an incubation time of z170 days and a 21 kDa
PrPSc (Prusiner, 1997). The experimental data demon-
strate that MHu2MPrPSc can exist in two different confor-
mations based on the sizes of the protease-resistant
fragments; yet, the amino acid sequence of MHu2MPrPSc
is invariant.
The results of our studies argue that PrPSc acts as a
template for the conversion of PrPC into nascent PrPSc .
Imparting the size of the protease-resistant fragment
of PrPSc through conformational templating provides a
mechanism for both the generation and propagation of
prion strains.
Interestingly, the protease-resistant fragment of PrPSc
after deglycosylation with an Mr of 19 kDa has been
found in a patient who died after developing a clinical
disease similar to FFI. Since both PrP alleles encoded
the wild-type sequence and a Met at position 129, we
Cell
344

labeled this case fatal sporadic insomnia (FSI). At au-

topsy, the spongiform degeneration, reactive astroglio-
sis, and PrPSc deposition were confined to the thalamus
(Mastrianni et al., 1997). These findings argue that the
clinicopathologic phenotype is determined by the con-
formation of PrPSc in accord with the results of the trans-
mission of human prions from patients with FFI to Tg
mice (Telling et al., 1996).
Mechanism of Selective Neuronal Targeting?
In addition to incubation times, neuropathologic profiles
of spongiform change have been used to characterize
prion strains (Fraser and Dickinson, 1968). However,
recent studies with PrP transgenes argue that such pro-
files are not an intrinsic feature of strains (Carp et al.,
1997; DeArmond et al., 1997). The mechanism by which
prion strains modify the pattern of spongiform degener-
ation was perplexing since earlier investigations had
shown that PrPSc deposition precedes neuronal vacuola-
tion and reactive gliosis (DeArmond et al., 1987). When
FFI prions were inoculated into Tg(MHu2M) mice, PrPSc
was confined largely to the thalamus (Figure 4A) as is the
case for FFI in humans (Telling et al., 1996). In contrast,
fCJD(E200K) prions inoculated into Tg(MHu2M) mice
produced widespread deposition of PrPSc throughout
the cortical mantel and many of the deep structures of
the CNS (Figure 4B) as is seen in fCJD(E200K) of hu-
mans. To examine whether the diverse patterns of PrPSc
deposition are influenced by Asn-linked glycosylation
of PrPC, we constructed Tg mice expressing PrPs mu-
tated at one or both of the Asn-linked glycosylation
consensus sites (DeArmond et al., 1997). These muta-
tions resulted in aberrant neuroanatomic topologies of
PrPC within the CNS, whereas pathologic point muta-
tions adjacent to the consensus sites did not alter the
distribution of PrPC. Tg mice with mutation of the second
PrP glycosylation site exhibited prion incubation times
of .500 days and unusual patterns of PrPSc deposition.
These findings raise the possibility that glycosylation
can modify the conformation of PrP and affect either
the turnover of PrPC or the clearance of PrPSc. Regional Figure 4. Regional Distribution of PrP Sc Deposition in
differences in the rate of deposition or clearance would Tg(MHu2M)Prnp0/0 Mice Inoculated with Prions from Humans Who
Died of Inherited Prion Diseases
result in specific patterns of PrPSc accumulation.
Histoblot of PrP Sc deposition in a coronal section of a
Tg(MHu2M)Prnp0/0 mouse through the hippocampus and thalamus
Yeast and Other Prions (Telling et al., 1996).
Although prions were originally defined in the context (A) The Tg mouse was inoculated with brain extract prepared from
of an infectious mammalian pathogen (Prusiner, 1982), a patient who died of FFI.
it is now becoming widely accepted that prions are ele- (B) The Tg mouse was inoculated with extract from a patient with
ments that impart and propagate variability through mul- fCJD(E200K). Cryostat sections were mounted on nitrocellulose and
treated with proteinase K to eliminate PrP C (Taraboulos et al., 1992).
tiple conformers of a normal cellular protein. Such a
To enhance the antigenicity of PrPSc , the histoblots were exposed
mechanism must surely not be restricted to a single to 3 M guanidinium isothiocyanate before immunostaining using
class of transmissible pathogens. Indeed, it is likely that a-PrP 3F4 MAb (Kascsak et al., 1987).
the original definition will need to be extended to encom- (C) Labeled diagram of a coronal section of the hippocampus/thala-
pass other situations where a similar mechanism of in- mus region. NC, neocortex; Hp, hippocampus; Hb, habenula; Th,
formation transfer occurs. thalamus; vpl, ventral posterior lateral thalamic nucleus; Hy, hypo-
Two notable prion-like determinants, [URE3] and thalamus; Am, amygdala.
[PSI], have already been described in yeast and one in
another fungus denoted [Het-s* ] (Wickner, 1994; Cher-
noff et al., 1995; Coustou et al., 1997). Studies of candi- molecular chaperone Hsp104; however, no homolog of
date prion proteins in yeast may prove particularly help- Hsp104 has been found in mammals (Chernoff et al.,
ful in the dissection of some of the events that feature 1995). The NH2-terminal prion domains of Ure2p and
in PrPSc formation. Interestingly, different strains of yeast Sup35 that are responsible for the [URE3] and [PSI]
prions have been identified (Derkatch et al., 1996). Con- phenotypes in yeast have been identified. In contrast to
version to the prion-like [PSI] state in yeast requires the PrP, which is a GPI-anchored membrane protein, both
Review
345
Ure2p and Sup35 are cytosolic proteins (Wickner, 1997).
When the prion domains of these yeast proteins were
expressed in E. coli, the proteins were found to polymer-
ize into fibrils with properties similar to those of proteo-
lytically trimmed PrP and other amyloids (Paushkin et
al., 1997).
Whether prions explain some other examples of ac-
quired inheritance in lower organisms is unclear (Land-
man, 1991). For example, studies on the inheritance of
positional order and cellular handedness on the surface
of small organisms have demonstrated the epigenetic
nature of these phenomena but the mechanism remains
unclear (Frankel, 1990).
Therapeutic Approaches to Prion Diseases
It seems likely that it will be possible to design effective
therapeutics for prion diseases as our understanding of
prion propagation increases. Because people at risk for
inherited prion diseases can now be identified decades
before neurologic dysfunction is evident, the develop-
ment of an effective therapy for these fully penetrant
disorders is imperative. Although we have no way of
predicting the number of individuals who may develop
neurologic dysfunction from bovine prions in the future,
seeking an effective therapy now seems most prudent
(Prusiner, 1997). Interfering with the conversion of PrPC
into PrPSc seems to be the most attractive therapeutic
target. Reasonable strategies are either stabilizing the
structure of PrPC via the formation of a PrPC-drug com-
plex or modifying the action of protein X, which may
function as a molecular chaperone (Figure 2B). Whether
it is more efficacious to design a drug that binds to PrPC
at the protein X–binding site or one that mimics the
structure of PrPC with basic polymorphic residues that
seem to prevent scrapie and CJD remains to be deter-
mined (Kaneko et al., 1997b; Shibuya et al., 1998). Since
PrPSc formation seems limited to caveolae-like domains
(Gorodinsky and Harris, 1995; Taraboulos et al., 1995),
drugs designed to inhibit this process need not pene-
trate the cytosol of cells but they do need to be able to
enter the CNS. Alternatively, drugs that destabilize the
structure of PrPSc might also prove useful.
The production of domestic animals that do not repli-
cate prions may also be important with respect to pre-
venting prion disease. Sheep encoding the R/R polymor-
phism at position 171 seem to be resistant to scrapie
(Hunter et al., 1993; Westaway et al., 1994); presumably,
this was the genetic basis of James Parry’s scrapie
eradication program in Great Britain 30 years ago (Parry,
1962). A more effective approach using dominant nega-
tives for producing prion-resistant domestic animals,
including sheep and cattle, is probably the expression
of PrP transgenes encoding R171 as well as additional
basic residues at the protein X–binding site (Figure 2B)
(Kaneko et al., 1997b). Such an approach can be readily
evaluated in Tg mice, and once shown to be effective,
it could be instituted by artificial insemination of sperm
from males homozygous for the transgene. Less practi-
cal is the production of PrP-deficient cattle and sheep.
Although such animals would not be susceptible to prion
disease (Büeler et al., 1993; Prusiner et al., 1993), they
might suffer some deleterious effects from ablation of
the PrP gene (Collinge et al., 1994; Lledo et al., 1996;
Sakaguchi et al., 1996; Tobler et al., 1996).
Whether gene therapy for the human prion diseases
will prove feasible using the dominant negative ap-
proach described above for prion-resistant animals de-
pends on the availability of efficient vectors for delivery
of the transgene to the CNS.
Concluding Remarks
Although the study of prions has taken several unex-
pected directions over the past three decades, a novel
and fascinating story of prion biology is emerging. In-
vestigations of prions have elucidated a previously un-
known mechanism of disease in humans and animals.
While learning the details of the structures of PrPs and
deciphering the mechanism of PrPC transformation into
PrPSc will be important, the fundamental principles of
prion biology have become reasonably clear. Though
some investigators prefer to view the composition of
the infectious prion particle as unresolved (Manuelidis
and Fritch, 1996; Lasmézas et al., 1997; Chesebro,
1998), such a perspective denies an enlarging body of
data, none of which refutes the prion concept. Moreover,
the discovery of prion-like phenomena mediated by pro-
teins unrelated to PrP in yeast and other fungi serves
not only to strengthen the prion concept but also to
widen it.
The discovery that prion diseases in humans are
uniquely both genetic and infectious greatly strength-
ened and extended the prion concept. To date, 20 differ-
ent mutations in the human PrP gene, all resulting in
nonconservative substitutions, have been found either
to be linked genetically to or to segregate with the inher-
ited prion diseases (Figure 1B). Yet, the transmissible
prion particle is composed largely, if not exclusively, of
an abnormal isoform of the prion protein designated
PrPSc (Prusiner, 1991).
Aberrant PrP Metabolism
The hallmark of all prion diseases—whether sporadic,
dominantly inherited, or acquired by infection—is that
they involve the aberrant metabolism and resulting ac-
cumulation of the prion protein (Table 1) (Prusiner, 1991).
The conversion of PrPC into PrPSc involves a conforma-
tion change whereby the a-helical content diminishes
and the amount of b sheet increases (Pan et al., 1993).
These findings provide a reasonable mechanism to ex-
plain the conundrum presented by the three different
manifestations of prion disease.
Understanding how PrPC unfolds and refolds into
PrPSc will be of paramount importance in transferring
advances in the prion diseases to studies of other de-
generative illnesses. The mechanism by which PrPSc is
formed must involve a templating process whereby ex-
isting PrPSc directs the refolding of PrPC into a nascent
PrPSc with the same conformation. Not only will a knowl-
edge of PrPSc formation help in the rational design of
drugs that interrupt the pathogenesis of prion diseases,
but it may also open new approaches to deciphering
the causes of and to developing effective therapies
for the more common neurodegenerative diseases in-
cluding Alzheimer’s disease, Parkinson’s disease, and
amyotrophic lateral sclerosis (ALS). Indeed, the ex-
panding list of prion diseases and their novel modes of
Cell
346
transmission and pathogenesis (Table 1), as well as the
unprecedented mechanisms of prion propagation and
information transfer, indicate that much more attention
to these fatal disorders of protein conformation is ur-
gently needed.
But prions may have even wider implications than
those noted for the common neurodegenerative dis-
eases. If we think of prion diseases as disorders of
protein conformation and do not require the diseases
to be transmissible, then what we have learned from the
study of prions may reach far beyond these common
illnesses.
Conformational Diversity
The discovery that proteins may have multiple biologi-
cally active conformations may prove no less important
than the implications of prions for diseases. How many
different tertiary structures can PrPSc adopt? This query
not only addresses the issue of the limits of prion diver-
sity but also applies to proteins as they normally function
within the cell or act to affect homeostasis in multicellu-
lar organisms. The expanding list of chaperones that
assist in the folding and unfolding of proteins promises
much new knowledge about this process. For example,
it is now clear that proproteases can carry their own
chaperone activity where the pro portion of the protein
functions as a chaperone in cis to guide the folding of
the proteolytically active portion before it is cleaved
(Shinde et al., 1997). Such a mechanism might well fea-
ture in the maturation of polypeptide hormones. Interest-
ingly, mutation of the chaperone portion of prosubtilisin
resulted in the folding of a subtilisin protease with differ-
ent properties than the one folded by the wild-type chap-
erone. Such chaperones have also been shown to work
in trans (Shinde et al., 1997). Besides transient metabolic
regulation within the cell and hormonal regulation of
multicellular organisms, it is not unreasonable to sug-
gest that assembly of proteins into multimeric structures
such as intermediate filaments might be controlled at
least in part by alternative conformations of proteins.
Such regulation of multimeric protein assemblies might
occur in either the proteins that form the multimers or
the proteins that function to facilitate the assembly pro-
cess. Additionally, apoptosis during development and
throughout adult life might also be regulated at least in
part by alternative tertiary structures of proteins.
Future Studies
The wealth of data establishing the essential role of PrP
in the transmission of prions and the pathogenesis of
prion diseases has provoked consideration of how many
biological processes are controlled by changes in pro-
tein conformation. The extreme radiation-resistance of
the scrapie infectivity suggested that the pathogen
causing this disease and related illnesses would be dif-
ferent from viruses, viroids, and bacteria (Alper et al.,
1967). Indeed, an unprecedented mechanism of disease
has been revealed where an aberrant conformational
change in a protein is propagated. The future of this
emerging area of biology should prove even more inter-
esting and productive as many new discoveries emerge.
Acknowledgments
We thank G. Carlson, N. Nathanson, and J. Safar for carefully re-
viewing sections of this manuscript. This research was supported
by grants from the National Institute of Aging and the National Insti-
tute of Neurologic Diseases and Stroke of the National Institutes
of Health, International Human Frontiers of Science Program, and
American Health Assistance Foundation, as well as by gifts from
the Sherman Fairchild Foundation, Keck Foundation, G. Harold and
Leila Y. Mathers Foundation, Bernard Osher Foundation, John D.
French Foundation, and Centeon.
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