Structural disorder,
prions, amyloids and polyglutamine diseases
Peter Tompa
Institute of Enzymology
Hungarian Academy of Sciences
Budapest, Hungary
Amyloid diseases
Disease
Alzheimer’s disease
Primary systemic amyloidosis
Senile systemic amyloidosis
Diabetes type II
Hemodialysis-associated amyloidosis
Familial systemic amyloidosis
Huntingon’s disease
Parkinson’s disease
CJD, other prion diseases
Taupathies, Pick disease, FTDP-17
Protein/peptide
Aggregate
Amyloid diseases: “traditional” classification
systemic vs. tissue-specific
juvenile vs. adult or old
inherited vs. spontaneous
primary vs. secondary
protein vs. peptide
mass in kgs vs. almost negligible
(globular vs. IUP)
so what is common ???
Amyloid fibrils
• 10 nm
• straight
• stable
• tinctorial
properties (Congo
red)
• cross-b
Symptoms fall into two broad classes
Systemic cases
- organ failure (heart, liver, kidney)
Tissue-specific cases
- cognitive impairment (dementia, often
with psychiatric symptoms)
- loss of coordination of movement
- neurodegeneration
Amyloid diseases
Disease
Protein/peptide
Aggregate
Alzheimer’s disease
A
Senile plaq
Primary systemic amyloidosis
Ig light chain
Senile systemic amyloidosis
Transthyretin
Diabetes type II
Amylin
Hemodialysis-associated amyloidosis
2-microglobulin
Familial systemic amyloidosis
Lysozyme mutant
Huntingon’s disease
Huntingtin
Huntingtin inclusion
Parkinson’s disease
-synuclein
Lewy body
CJD, other prion diseases
PrPSc
Prion aggregate
Taupathies, Pick disease, FTDP-17
Tau protein
PHF, Pick-body
Amyloid diseases: modern classification
 protein misfolding diseases
1) Protein (AL, ATTR, ALys)
2) Cause (spontaneous, mutation, induced)
3) Mechanism (loss or gain of function)
Alzheimer’s disease
AD plaque
Neurofibrillary tangle (PHF)
Amyloid precursor protein (APP)
(TACE, ADAM10)
(PSEN)
„Lag-phase” and „seeding” (1D crystal growth)
„Seeding”
Exponential
growth
Long
incubation time
Chen et al. (2001) JMB 311, 173
Familial systemic amyloidosis:
Lysozyme mutants
D67H
I56T
Reduced stability of amyloidogenic mutants
Wild type
Ile56Thr
Asp67His
Booth et al. (1997) Nature 385, 787
123I-SAP
scintigraphy
liver
normal
kidney
Pepys
Huntington’s disease
(Huntingtin)
MATLEKLMKAFESLKSFQQQQQQQQQQQQQQQQQQQQQQQP
PPPPPPPPPPQLPQPPPQAQPLLPQPQPPPPPPPPPPGPAV
AEEPLHRPKKELSATKKDRVNHCLTICENIVAQSVRNSPEF
QKLLGIAMELFLLCSDDAESDVRMVADECLNKVIKALMDSN
LPRLQLELYKEIKKNG…
ATGGCGACCCTGGAAAAGCTGATGAAGGCCTTCGAGTCCCTCAA
GTCCTTCCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGC
AGCAGCAGCAGCAGCAGCAGCAGCAGCAACAGCCGCCACCGCCG
CCGCCGCCGCCGCCGCCTCCTCAGCTTCCTCAGCCGCCGCCGCA
GGCACAGCCGCTGCTGCCTCAGCCGCAGCCGCCCCCGCCGCCGC
CCCCGCCGCCACCCGGCCCGGCTGTGGCTGAGGAGCCGCTGCAC
CGACCAAAGAAAGAACTTTCAGCTACCAAGAAAGACC…
PolyQ expansion: polymorphisms
Wells (1996) JBC 271, 2875
Huntingtin inclusions in
neuronal nuclei
Perutz (1999) TiBS 24, 58
Cause of disease?
Loss of function
Gain of function
Anticipation in polyQ-disease inheritance
Age of onset, DRPLA
- dynamic mutation, mutable mutation -
CAG repeat units
Tsuji (1997) Int. Med. 36, 3
Prion diseases (TSE)
HUMAN
ANIMAL
SCRAPIE
sheep
kuru
BSE
bovine
CJD (Creutzfeldt-Jakob)
TME
mink
GSS (Gerstmann,Straussler,
CWD
deer
FSE
cat
Sheinker)
FFI
• rapid cognitive impairment (dementia)
• movement disorders
• spongiform degeneration
Chronology
• XVIII c.
• 1920
• 1939
• 1954
• 1959
• 1959
• 1966
• 1982
• 1986
• 1997
scrapie
CJD (heritable)
scrapie transmissible
scrapie: „slow virus”
kuru resembles CJD
kuru resembles scrapie
kuru  chimpanzee transmission GAJDUSEK
„prion” Prusiner
BSE (first case)
Nobel prize PRUSINER
Ancient scrapie?
disease
= like rash
`
،
flee
Wickner (2005) Science 309, 864
Chronology
• XVIII c.
• 1920
• 1939
• 1954
• 1959
• 1959
• 1966
• 1982
• 1986
• 1997
scrapie
CJD (heritable)
scrapie transmissible
scrapie: „slow virus”
kuru resembles CJD
kuru resembles scrapie
kuru  chimpanzee transmission GAJDUSEK
„prion” Prusiner
BSE (first case)
Nobel prize PRUSINER
Stanley B. Prusiner
• strange pathogen (resistance to UV, heat etc…)
• purification
• transmission to mouse (incubation time 150-300 days)
• 1975-77: transmission to hamster (70 days)
infected
proteinase K
C
SC
PrP
PrP
PrPsen PrP27-30
PrPres
1
232
Infectious protein ?
• no DNA
• PrPsc and infectivity purify together
• properties of PrPsc match those of prion
• PrP: encoded by the host
• inherited forms: mutations of PrP gene
1982
proteinaceous
infectious
PRION
Patholopgical prion: structure of PrPC
GPI
*
**
(PHGGGWGQ)5
*
*
*
*
* P102L
* P107L
*
A127GAAA*AGAVVGGLGG133
Amyloid: mad-cow disease
Extension of the prion concept:
physiological prions
Two yeast genetic element [URE3], [PSI+]
• dominant, non-Mendelian inheritance (meiois)
• non-chromosomal (cytoplasmic)
• metastable (curable)
• selective advantage ?
Sup35p (translation release factor 3, eRF3)
normal Sup35p = [psi-]
prion Sup35p = [PSI+]
Suppression
of nonsense
mutations
Sup35p: eukaryotic translation release factor3
MSNPQDQLSNDLANASISGDQSKQPQQQQPQQQQPY
FNPNQAQAFVPTGGYQQFQPQQQQQYGGYQQNYTQY
QAGGYQQNYNNRGGYQQNYNNRGGYQQNYNNRGGYQ
QQQQQQYQAYNPNQQYGGYQAYNPQQQQQQQTQSQG
MSLADFQKQKAEQQASLNKPAVKKTLKLASSSGIKL
ANATKKVDTAKPAASKEASPAPKDEEASAEPEAKKE
STPVPASSSPAPAAADSTPAPVKKESTPTPSVASKS
APVSASASVVTADALAKEQEDEVDEEVVKDMFGGKD
HVSIIFMGHVDA........
Prion (amyloid) form of Sup35 promotes
translation read-through
Sup35: disorder and modularity
Sup35: disorder and modularity
„Lag-phase” and „seeding” (1D crystal growth)
„Seeding”
Exponential
growth
Long
incubation time
Chen et al. (2001) JMB 311, 173
Prion infection: „cross-seeding”
„Cross-seeding”
Exponential
growth
Long
incubation time
Chen et al. (2001) JMB 311, 173
Extension of prion concept:
prions and memory?
Hippocampus and memory
Aplysia californica
LTF
GSW reflex
habituation,
sensitisation
Eric Kandel
Aplysia neuronal CPEB is involved in LTF
5 x 5-HT
Si et al. (2004) Cell 115, 893
Aplysia neuronal CPEB is a prion
Si et al. (2004) Cell 115, 879
The structure of amyloid(ogenic) proteins
Needs to be addressed:
- structure of amyloidogenic protein
- structure of intermediate
- structure of amyloid itself
Structure of amyloidogenic proteins
Globular:
lysoyzme
transthyretin (TTR)
insulin
b2-microglobulin
IDP:
-synuclein
tau protein
polyQ regions
prion domains
Structure: lysozyme
D67H
I56T
Structure: polyQ
Structural ensemble of -synuclein
(NMR paramagnetic relaxation
enhancement)
Dedmon et al. (2005) JACS 127, 476
Structure of amyloidogenic proteins
Globular:
lysoyzme
transthyretin (TTR)
insulin
b2-microglobulin
IDP:
-synuclein
tau protein
polyQ regions
prion domains
Structure of amyloidogenic proteins
Globular:
partial unfolding
lysoyzme
transthyretin (TTR)
insulin
b2-microglobulin
IDP:
partial folding
-synuclein
tau protein
polyQ regions
prion domains
Structure of the intermediate ?
temp.
temp.
Partially ordered amyloid precursors
Uversky and Fink (2005) BBA 1698, 131
The common denominator: polyproline II helix?
SH3-PPII
Wikipedia
PPII in a-synuclein (ROA)
Syme (2002) EJB 269, 148
57°C pH 2.0
Structure of amyloid: cryo-EM
A reasonable analogy: the Leu zipper
GCN-4 bZip
Polar zipper (vs. Leu zipper)
Perutz (1994) Prot. Sci. 3, 1629
Structure of Sup35 prion peptide (steric zipper)
DSNQGNNQQNYQQY
SQNGNQQQGNNRYQ
GYQAYNAQAQPAGG
YYQNYQGYSGYQQG
GYQQYNPDAGYQQQ
YNPQGGYQQYNPQG
GYQQQFNPQ
Nelson et al. (2005) Nature 435, 773
Structure of A(beta) (1-40) protofilament
Luhrs (2005) PNAS 102, 16248
Structural model of the CA150.WW2 protofilament
Ferguson (2006) PNAS 103, 162
Points of interference
Dobson (2004) Science 304, 1259