PRIONS

Definition
• PRION: Infectious protein
particles thought to be
responsible for a group of
transmissible and/or inherited
neurodegenerative diseases.
• "Prion" is short for
"Proteinaceous Infectious
particle"
 Structure
Prions are infectious agents composed
exclusively of a single sialoglycoprotein
called PrP 27-30.
They contain no nucleic acid.
PrP 27-30 has a mass of 27,000 -
30,000 daltons and is composed of 145
amino acids with glycosylation at or
near amino acids 181 and 197.
This protein polymerizes into rods
possessing the ultrastructural and
histochemical characteristics of
amyloid.
 Evidence suggests that a prion is a
modified form of a normal cellular
protein known as PrPc (for cellular), a
single copy gene.
 This protein is found predominantly on
the surface of neurons attached by a
glycoinositol phospholipid anchor,
and is protease sensitive. Thought to
be involved in synaptic function.
 The modified form of PrPc which may cause disease
i.e.
the prion is known as PrPsc (for scrapie) which is
relatively resistant to proteases and
accumulates in
cytoplasmic vesicles of diseased individuals.
 It has been proposed that PrPsc when introduced
into a normal cell causes the conversion of PrPc into
PrPsc.
Process is unknown but it could involve a chemical or
conformational modification.
 Replication
The prion is a product of a human gene, termed the
PrP gene, found on chromosome 20.
This gene contains two exons separated by a single
intron.
Exon I and Exon II are transcribed and the two RNAs
ligated into a single mRNA.
This mRNA contains an open reading frame (ORF) or
protein coding region which is translated into the PrP
protein.
The PrP protein is a precursor of the prion protein. It
is termed PrP 33-35.
The PrP 33-35 undergoes several post-
translational events to
become the prion protein (PrP 27-30):
1. Glycosylation - at two sites.
2. Formation of a disulfide bond between two
cysteine residues.
3. Removal of the N-terminal signal peptide.
4. Removal of the C-terminal hydrophobic
segment.
5. Addition of a phosphatidylinositol glycolipid at
the Cterminal.
6. Removal of the N-terminal first 57 amino
acids.
Pathologies induced by
prions
All diseases known to be of prion
etiology, in animals and
humans, are neurodegenerative
diseases. In the human this
includes:
•Creutzfeldt-Jakob disease (CJD)
•Fatal Familial Insomnia
•Gerstmann-Straussler syndrome
•Kuru
The pathological and clinical signs of
these diseases suggest
that they are closely related.
In fact they may be variants of the
same disorder.
All pathological features are confined to
the central nervous
system.
The prion protein accumulates
selectively and abnormally in
CNS nerve cells during the course of
the disease.
PrP 27-30 accumulates within
the neuronal system where it
causes:
1. Astrocyte gliosis (an increase
in the number of
astrocytes).
2. Depletion of dendritic spines
in neurons.
3. Formation of numerous
vacuoles in the cerebellar cortex
(spongiform encephalopathy).
4. Amyloidosis - deposition of amyloid in the
cerebellar сortex, thalamus, brain stem and in the
lumen of blood vessels within the brain.
These amyloid plaques consist of discrete
eosinophilic glassy-appearing masses, often
having radiating amyloid fibrils at their periphery.
The plaques are primarily subependymal, subpial
and perivascular.
Note that the pathology does NOT
include any signs of
inflammation or fever.
This is evidence that the immune system
does not respond to
the prion protein.
Since the prion protein is derived from
self this is what you
would expect.
These pathologies give rise to the
clinical symptomology
seen in these patients.
These are:
1. A long incubation period (several years) which has
given
rise to the term "slow infection."
2. Loss of muscle coordination which leads to a difficulty
in walking, indicating a functional disorder of the
cerebellum.
3. Dementia characterized initially by loss of memory,
diminished intellect and poor judgment.
4. Progressive insomnia characterized by a marked
reduction or loss of the slow-wave and rapid-eye-
movement phases.
Transmission

Spread of the disease is via

horizontal transmission, i.e.,
transmission from one person to
another, either directly or
by ingestion of contaminated
meat.
 Diagnosis
In the past, diagnosis of prion disease was
made through examination of brain biopsies
taken from patients in advanced stages of the
disease or, more commonly, after
they had died.
In January of 1999 it was found that the prion
protein accumulated in the tonsils and could be
detected by an immunofluorescence test on
tonsilar biopsies.
A second test was simultaneously
developed which was
based on a Western blot.
Later that year a third test was
developed that had the high
sensitivity necessary to detect the
prion protein in blood.
This test is based on capillary
electrophoresis with laserinduced
fluorescence. It detects as little as
10-18 mole.
Prion diseases are often called spongiform
encephalopathies
because of the post mortem appearance of the
brain with
large vacuoles in the cortex and cerebellum.
Probably most mammalian species develop these
diseases.
Specific examples include:
•Scrapie: sheep
•TME (transmissible mink encephalopathy): mink
•CWD (chronic wasting disease): muledeer, elk
•BSE (bovine spongiform encephalopathy): cows
Humans are also susceptible
to several prion diseases:
•CJD: Creutzfeld-Jacob Disease
•GSS: Gerstmann-Straussler-
Scheinker syndrome
•FFI: Fatal familial Insomnia
•Kuru
•Alpers Syndrome
•The incidence of sporadic CJD is about 1
per million per year.
•GSS occurs at about 2% of the rate of
CJD.
•It is estimated that 1 in 10,000 people are
infected with CJD
at the time of death.
These figures are likely to be
underestimates since prion
diseases may be misdiagnosed as other
neurological
disorders.
•The diseases are characterized by loss of
motor control, dementia, paralysis wasting
and eventually death,
typically following pneumonia.
• Fatal Familial Insomnia presents with an
untreatable
insomnia and dysautonomia. Details of
pathogenesis are largely unknown.
•Visible results at post-mortem are non-
inflammatory lesions, vacuoles, amyloid
protein deposits and
astrogliosis.
•GSS is distinct from CJD, it
occurs typically in the 4th-5th
decade, characterized by
cerebellar ataxia and
concomitant
motor problems, and less
common dementia.
•(Originally thought to be
familial, but now known to occur
sporadically as well).
•CJD typically occurs a decade later has
cerebral involvement so dementia is more
common and patient seldom survives a
year (originally thought to be sporadic, but
now known to be familial as well).
•FFI pathology is characterized by severe
selective atrophy of the thalamus.
Alpers syndrome is the name given to
prion diseases in infants.
• Kuru is the condition which first
brought prion diseases to
prominence in the 1950s. Found in
geographically isolated tribes in the New
Guinea.
• Established that ingesting brain tissue
of dead relatives for
religious reasons was likely to be the
route of transmission.
They ground up the brain into a pale grey
soup, heated it and
ate it.
• Clinically, the disease resembles CJD.
Other tribes in the vicinity with same
religious habit did not develop the disease.
It is speculated that at some point in the
past a tribe member developed CJD, and as
brain tissue is highly infectious this allowed
the disease to spread. Afflicted tribes were
encouraged
not to ingest brain tissue and the incidence
of disease rapidly
declined and is now almost unknown.
Humans might be infected by
prions in 2 ways:
1. Acquired infection (diet and following
medical procedures
such as surgery, growth hormone
injections, corneal
transplants) i.e. infectious agent implicated.
2. Apparent hereditary Mendelian
transmission - where it is
an autosomal and dominant trait. This is not
consistent with an infectious agent.
• This is one of the features that single
out prion diseases for particular
attention.
• They are both infectious and
hereditary diseases.
• They are also sporadic, in the sense
that there are also cases in which there
is no known risk factor although it
seems likely that infection was
acquired in one of the two ways listed
above.
•Scrapie was the first example of this type of
disease to be noticed and has been known
about for many hundreds of years.
•There are two possible methods of
transmission in sheep:
1. Infection of pasture with placental tissue
carrying the agent followed by ingestion,or
direct sheep-lamb transmission i.e. an
acquired infection.
2. Parry showed considerable foresight by
suggesting that it is not normally an infectious
disease at all but a genetic disorder. He
further suggested that selective breeding
would get rid of the disease.
 Treatment
•PrPc overexpression facilitates the development of
prion diseases.
It may therefore follow that agents which reduce PrPc
expression will delay the onset of prion diseases.
One can speculate that chemicals which bind to and
stabilize the PrPc conformation may be beneficial.
Similarly agents destabilizing the PrPsc conformation
may be effective. In this regard several vaccines to
Alzheimers amyloid plaques are in clinical trials.
Agents which interfere with the putative PrPc-PrPsc
interaction might similarly be effective.
•A number of reagents showing
affinity for amyloid proteins
are known e.g congo red.
As our knowledge of the
structure of PrPc increases, the
chances of rationally deducing
effective therapeutics based
on these ideas increases.
•Finally we have seen that PrPc expression is
required for pathology.
Chemicals affecting the endocytosis, exocytosis,
intracellular trafficking and degradation of
proteins and in particular PrPc
may also be effective.
Amphotericin for instance is reported to delay
prion disease in hamsters (although it
apparently has little effect in
humans).
• References:
Prions.http://www.cartage.org.lb/en/themes/
Sciences/LifeScience/GeneralBiology/Micr
obiology/PrionsViroids/Prions/Prions.htm