PRIONS

THE
INFECTIOUS PROTEINS
Paras Yadav*, Jaspreet Singh Arora*, Sachinandan De*, Tirtha
Kumar Datta*, Surender Lal Goswami*, Aarti Bhardwaj$,
Shalini Jain# and Hariom Yadav#
*Animal Biotechnology, #Animal Biochemistry Division, National Dairy
Research Institute, Karnal-132001, Haryana, India
$

Meerut Institute of Engeenering and Technology, Meerut, U.P., India

INTRODUCTION

Stanley B. Prusiner coined the term proin from Proteinaceous infective particle
and changed to prion to sound it rhythmic.

Prion diseases were caused by misfolded proteins.

Elucidated the gene and mechanism by which wild type protein bring about the
clinical disease.

Prion Diseases
Human

Kuru

Animal

Scrapie

Fatal Familial Insomnia (FFI)

Bovine Spongiform
Encephalopathy (BSE)

Creutzfeldt-Jakob disease
(CJD)

Chronic Wasting Disease

(CWD)

Classification of prion diseases

Infectious/Exogenous
e.g., Kuru, BSE (mad cow disease), Scrapie
Spread by
Consumption of infected material.
Transfusion.

Sporadic
Familial/Hereditary
Due to autosomal dominant mutation of PrP.

Differences between cellular and scrapie protein

PrPC

PrPSC
Solubility

Soluble

Non soluble

Structure
Alpha-helical

Beta-sheeted

Multimerisation state
Monomeric

Multimeric

Infectivity
Non infectious

Infectious

Susceptibility to Proteinase K
Susceptible

Resistant

Steps in the biosynthesis of PrP

Post-translational processing of PrP

Cellular trafficking and cleavage of PrP

ER

Endosome

Golgi

Mechanism of Internalization of PrP

Hypothetical model for a PrP receptor

Model for the function of

LRP- LR as the receptor for PrP
BDII (aa 53-93)

BDI
(aa 144-179)

PrP

HSPG
Dependent
binding domain
Direct binding
domain (aa
161-179)
LRP/LR

GPI

Heparan sulfate chain

(HS)

Proteogl
ycan
moiety

Sulfated domains

HSPG

Cell Culture Systems for Prion Propagation

Sequence of prion protein

Functions of Prion protein

Antioxidative

PrPC + Cu (Copper)

Antioxidant activity

Resistance to oxidative stress

Prevent neuronal dysfunction

(Brown et al., 2002)

Other functions

Models for the conversion of PrP to PrP

sc

Sc

Time taken for Transformation of mutant PrP to a PrP state

Synthesis of
Mutant PrPc

<10 min

PIPLCresistant

Endoplasmic Reticulum

Detergent
insoluble

Plasma membrane/

BFA
180C

0.5-1 hr

Endocytic Pathway

1-6 hr

Proteaseresistant

Effect of conformation of PrP on Pro K

Model of the cellular pathways

Sc
involved in generation of PrP

Proposed model of PrP

aggregation and induction of CtmPrP

Pathogenesis

sc

Mechanism of PrP induced apoptosis

What are Calpains?

They generate a C-terminal fragment(C2) which has molecular
weight of 27-30 kDs.

Increase in intracellular levels of Calcium increase

production of terminal fragments .

Calpastatin prevents production of C2.

Inhibitors of lysosomal proteases has no effect on C2
production.

Telling et al.,2004

Role of Caspases
It was proposed that prion-associated toxicity involves altered
trafficking of PrPc.

Inhibition of ubiquitin-proteasome system(UPS).

Deposition of aggresomes of PrPSc in nerve cells.
Induction of Apoptosis with activation of Caspase 3 and
Caspase 8.

Complete molecular basis for neuronal death is not known.

Aggregates of over expressed PrPc does not cause cell death.
Tabrizi et al., 2005

Factors Responsible for Prion Propagation

The AGAAAAGA Palindrome in Prion Generation

Norstrom & Mastrianni, 2005

Factors Responsible for Prion Propagation cont

PrPc association with lipid rafts in the early secretory pathway.

CreutzfeldtJakob disease (CJD) in Libyan Jews, linked to the E200K

mutation in PRNP (E200KCJD).

Model for chaperone-supervised PrP conversion

E.g. Hsp70, GroEL

Factors that prevent Prion Replication

Phospholipase A2 Inhibitors prevent prion replication.
Platelet-activating Factor Antagonists also inhibits prion replication.
Bate et al.,2004

Drugs which share a N-benzylidene-benzohydrazide core structure.

Bertsch et al.,2005

Trimethylamine N-oxide (TMAO), can prevent formation of PrPSc.

Bennion, 2004

Gross and Microscopic Changes

Gross changes
Grossly there is Cortical atrophy and
may also be present.

ventricular

dilatation

Microscopic changes

Scrapie

BSE

Kuru

CJD

Other microscopic changes

Gliosis within plaques.

Loss of oligodendrocytes within plaques.

Axons usually remain intact in plaques.

Both CD4+ and CD8+ lymphocytes are present in active

lesions.
(Kretzschmar et al.,1996, Wilesmith et al., 1997).

Diagnosis
1.
2.
3.
4.
6.

Diagnosis can be made by:

Clinical signs and Symptoms.
Detection of Scrapie
Associated fibrils.
Detection of Abnormal Prion
protein (PrPsc) by Western blotting.
Scrapie Associated
fibrils.
Two dimensional Gel Electrophoresis.
5.
Imunodiagnosis of Prion disease.
Bioassay in Mice.

Mechanism of plaque formation

PrPC

PrPSC

PrPC

PrPsc fibrils

Plaque

Conclusion
Molecular hallmark of the disorder is the accumulation of abnormal prion
protein(PrPSc).

Physiological functions of cellular prion protein (PrPc) is not clear.

Identity of intracellular compartment where PrPc to PrPSc occurs is not
established.

Prion peptide of 106-126 residues is found to be neurotoxic.