Structural disorder,Structural disorder,

prions, prions, amamyyloidloids and s and polpolyyglutaminglutaminee diseasesdiseases

Institute of EnzymologyHungarian Academy of Sciences

Budapest, Hungary

Peter Tompa

AmAmyyloid loid diseasesdiseases

Disease Protein/peptide Aggregate

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

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

1) Protein (AL, ATTR, ALys)

2) Cause (spontaneous, mutation, induced)

3) Mechanism (loss or gain of function)

Amyloid diseases: modern classification

protein misfolding diseases

AD plaque Neurofibrillary tangle (PHF)

Alzheimer’s disease

Amyloid precursor protein (APP)

(TACE, ADAM10)

(PSEN)

„Lag-phase” and „seeding” (1D crystal growth)

Chen et al. (2001) JMB 311, 173

Long

incubation time

Exponential growth

„Seeding”

Familial systemic amyloidosis:

LLysysozozyymmee mut mutantsants

I56T

D67H

Reduced stability of amyloidogenic mutants

Wild type

Ile56Thr

Asp67His

Booth et al. (1997) Nature 385, 787

normal

kidney

liver

123I-SAP scintigraphy

Pepys

Huntington’s disease (Huntingtin)

MATLEKLMKAFESLKSFQQQQQQQQQQQQQQQQQQQQQQQPPPPPPPPPPPQLPQPPPQAQPLLPQPQPPPPPPPPPPGPAVAEEPLHRPKKELSATKKDRVNHCLTICENIVAQSVRNSPEFQKLLGIAMELFLLCSDDAESDVRMVADECLNKVIKALMDSNLPRLQLELYKEIKKNG…

ATGGCGACCCTGGAAAAGCTGATGAAGGCCTTCGAGTCCCTCAAGTCCTTCCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAACAGCCGCCACCGCCGCCGCCGCCGCCGCCGCCTCCTCAGCTTCCTCAGCCGCCGCCGCAGGCACAGCCGCTGCTGCCTCAGCCGCAGCCGCCCCCGCCGCCGCCCCCGCCGCCACCCGGCCCGGCTGTGGCTGAGGAGCCGCTGCACCGACCAAAGAAAGAACTTTCAGCTACCAAGAAAGACC…

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

- dynamic mutation, mutable mutation -

Tsuji (1997) Int. Med. 36, 3

Ag

e o

f on

set,

DR

PLA

CAG repeat units

Prion diseases (TSE)

ANIMALANIMAL

SCRAPIESCRAPIE sheepsheep

BSEBSE bovinebovine

TMETME minkmink

CWDCWD deerdeer

FSEFSE catcat

HUMANHUMAN

kurukuru

CJD (Creutzfeldt-Jakob)CJD (Creutzfeldt-Jakob)

GSS (Gerstmann,Straussler, GSS (Gerstmann,Straussler,

Sheinker)Sheinker)

FFIFFI

• rapid cognitive impairment (dementia)• movement disorders• spongiform degeneration

Chronology

• XVIIIXVIII c. c. scrapiescrapie• 19201920 CJD (CJD (heritableheritable))• 19391939 scrapie scrapie transmissibletransmissible• 19541954 scrapie: „slow virus”scrapie: „slow virus”• 19591959 kuru kuru resemblesresembles CJD CJD• 19591959 kuru kuru resemblesresembles scrapie scrapie• 19661966 kuru kuru c chimpanzeehimpanzee transmissi transmissionon GAJDUSEKGAJDUSEK• 19821982 „prion” Prusiner„prion” Prusiner• 19861986 BSE (BSE (first case)first case)• 19971997 NobelNobel prize prize PRUSINERPRUSINER

Ancient scrapie?

Wickner (2005) Science 309, 864

fleedisease = like rash

`

،

Chronology

• XVIIIXVIII c. c. scrapiescrapie• 19201920 CJD (CJD (heritableheritable))• 19391939 scrapie scrapie transmissibletransmissible• 19541954 scrapie: „slow virus”scrapie: „slow virus”• 19591959 kuru kuru resemblesresembles CJD CJD• 19591959 kuru kuru resemblesresembles scrapie scrapie• 19661966 kuru kuru c chimpanzeehimpanzee transmissi transmissionon GAJDUSEKGAJDUSEK• 19821982 „prion” Prusiner„prion” Prusiner• 19861986 BSE (BSE (first case)first case)• 19971997 NobelNobel prize prize PRUSINERPRUSINER

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)

P rPC

P rP sen P rP 2 7-3 0P rP res

P rPS C

1 2 32

infectedinfected

proteinproteinasease K K

Infectious protein ?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

1982proteinaceous infectious

PRION

Patholopgical prion: structure of PrPC

(PHGGGWGQ)5

A127GAAA*AGAVVGGLGG133

GPI

***

*

*

**

*

*P107L* P102L

Amyloid: mad-cow disease

Extension of theExtension of the prion prion conceptconcept::

physiologicalphysiological prion prionss

Two yeastTwo yeast geneti genetic elementc element [[URE3URE3]], , [[PSI+PSI+]]

• domindominantant, n, nonon--Mendelian inheritanceMendelian inheritance (mei(meioois)is)• non-chnon-chromosromosomalomal (c (cyytoplatoplasmicsmic))• metastabmetastablele (curable) (curable)• sselective advantage ?elective advantage ?

normnormalal Sup35p = Sup35p = [[psipsi--]]

prion Sup35p = prion Sup35p = [[PSIPSI++]]

Sup35p (translation release factor 3, eRF3)

Suppression Suppression of nof nonsensonsensee mutmutationsations

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” Lag-phase” andand „seeding” „seeding” (1D crystal (1D crystal growth)growth)

Chen et al. (2001) JMB 311, 173

Long

incubation time

Exponential growth

„Seeding”

Prion infection:Prion infection: „ „cross-cross-seeding”seeding”

Chen et al. (2001) JMB 311, 173

Long

incubation time

Exponential growth

„Cross-seeding”

Extension of prion concept:

prions and memory?

Hippocampus and memory

Aplysia californica

habituation, sensitisation

LTF GSW reflex

Eric Kandel

Aplysia neuronal CPEB is involved in LTF

5 x 5-HT

Si et al. (2004) Cell 115, 893

Si et al. (2004) Cell 115, 879

Aplysia neuronal CPEB is a prion

The structure of amyloid(ogenic) proteins

Needs to be addressed:Needs to be addressed:

- structure of amyloidogenic - structure of amyloidogenic proteinprotein

- structure of intermediate- structure of intermediate

- structure of amyloid itself- structure of amyloid itself

Structure of amyloidogenic proteins

Globular:lysoyzme

transthyretin (TTR)

insulin

b2-microglobulin

IDP:-synuclein

tau protein

polyQ regions

prion domains

Structure: lyslysozozyymmee

I56T

D67H

Structure: pStructure: polyQolyQ

Dedmon et al. (2005) JACS 127, 476

Structural ensemble of -synuclein

(NMR paramagnetic relaxation enhancement)

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

temp.temp.

Structure of the intermediate ?Structure of the intermediate ?

Partially ordered amyloid precursors

Uversky and Fink (2005) BBA 1698, 131

Wikipedia

SH3-PPII

The common denominator: polyproline II helix?The common denominator: polyproline II helix?

PPII in a-synuclein PPII in a-synuclein (ROA)(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

Nelson et al. (2005) Nature 435, 773

Structure of Sup35 prion peptide (steric zipper)

DSNQGNNQQNYQQYSQNGNQQQGNNRYQGYQAYNAQAQPAGGYYQNYQGYSGYQQGGYQQYNPDAGYQQQYNPQGGYQQYNPQGGYQQQFNPQ

Structure of A(beta) (1-40) protofilament

Luhrs (2005) PNAS 102, 16248

Structural model of the CA150.WW2 Structural model of the CA150.WW2 protofilamentprotofilament

Ferguson (2006) PNAS 103, 162

PPooints of interferenceints of interference

Dobson (2004) Science 304, 1259