Parkinson’s Disease Mechanisms

HGNC Approved Symbol Mutation Gene Locus Inheritance Disease onset

SNCA Park 1/Park 4 α-synuclein 4q21.3-q22 Dominant Early

PARK2 Park 2 Parkin 6q25.2-q27 Recessive Juvenile; Early

PARK3 Park 3 ? 2p13 Dominant Late

UCHL1 Park 5 UCHL1 4p13 Dominant Late

PINK1 Park 6 PINK1 1p36.12 Recessive Early

PARK7 Park 7 DJ-1 1p36.23 Recessive Early

LRRK2 Park 8 LRRK2 12q12 Dominant Late

ATP13A2 Park 9 ATP13A2 1p36 Recessive Juvenile KRS, Early PD

PARK10 Park 10 AAOPD 1p32 Unclear Late

PARK11PARK12

Park 11 GIGYF2 2q36-q37 Recessive Early

HTRA2

Park 12 ? Xq21-q25 Unclear Late

PLA2G6

Park 13 Omi/HTRA2 2p13.1 Unclear Late

FBXO7

Park 14 PLA2G6 22q13.1 Recessive Juvenile LDOPA-responsive dystonia Parkinsonism

PARK16

Park 15 FBX07 22q11.2-qter

Recessive Early parkinsonian- pyramidal syn.

VPS35

Park 16 ? 1q32 Unclear Unclear

EIF4G1

Park 17 VSP35 16q12 Unclear Unclear

Park 18 EIF4G1 3q27-qter Unclear Unclear

PD-related gene mutations

α-Synuclein

Protofibrils

Fibrils

Neurodegeneration

Decreased Neurotoxicity

ProteinDegradation

PinkDegraded

ES ProteosomalDegradation Protective

Cell Death

Autophagy

Clinical Onset20 year disease stage20 year prodrome

HyposmiaConstipation

Bladder DisorderSYMPTOMS

HOEN / YAHR STAGE

PATHOLOGY

Braak Stage 1 2 3 4 5 6

Enteric Plexus;Olfactory Bulb;

CNX; Dorsal IX/X MN

Sleep Disorder,Obesity

Depression

Coerulus,Caudal Raphe

and Magnocellular RF

Unilateral Tremor,Rigidity, Bradykinesia

Substantia Nigra;Amygdala

(CN); Meynert’s Nucleus; PPN

BilateralDisease

Temporal Lobe: TEC;CA-2 Plexus; Intralaminar

Thalamic Nuclei; Signicant loss of melano-neurons

Poor Balance

Prefrontal Cortex: Tertiary Sensory Association Areas

Falls,Dependency,

Cognitive Decline

Chair/Bed Bound,Dementia

Secondary, then Primary Motor and Sensory Areas

Autophagy

Parkinson’s Disease Timeline

Dopamine Production Pathway

α-Synuclein Structure

Inflammation

Toxicity

Excreted

Toxicity

Excessive ROS Oxidative Stress

Excessive destruction or Accumulation of defective

mitochondria

Interaction with microtubule network

Synaptic vesicle formation

Amphipathic Region

Helix 1

Point mutations A30P E46K A53T

Helix 2

NAC Region Acidic Tail

Calcium homeostasis regulation

Catecholamine metabolismin DAergic neurons

Increased Mitophagy

Parkin

α-Methyl-paratyrosine

CarbidopaBenserazide

Pargyline

Tyrosine hydroxylase (TH)(tetrahydrobiopterin)

L-aromatic amino aciddecarboxylase (AADC)(Pyridoxal phosphate)

Monoamine oxidase (MAO)

Aldehyde dehydrogenase (ALDH)

Catechol-o-methyltransferase (COMT)

Tyrosine

MPTPMAO-B

DAT

D2 Autoreceptor

Rotenone

BLOO

D VE

SSEL

ASTROCYTE

POSTSYNAPTIC CELL

MPP+

Dihydroxy-phenylalanine(L-DOPA)

Dihydroxyphenyl-acetaldehyde

(DOPAL)

Dihydroxyphenyl-acetic acid(DOPAG)

Homovanillicacid (HVA)

OxidativeStress

O2-

Tetrabenazine

UCHL1

Caspases 1,8

Clioquinol

α-Sp22

Synphillin-1

SIAH

HSP70

CHIP

Ubc6/7

CDCRel-1/Septin5

EPRS

Cyclin E

Ataxin

Proteasome

Tubulins

Synaptotagmin XIUbCH7/8

UBA1 AMP + PPiATP

cullin1

Ligase

Hydr

olas

e

FBXw7

PAELRPAELR

PINK1p64

DA Oxidation

O-glycosylation

Glycation

Accumulation ofunfolded PAELR

Increased inhibitionof DA release

Mutant decreases hydrolase activity

Cx Ie-

MAO

NF-κB

NADH

NAD+

DA

DOPAL + H2O2

α-KG

Succiny-CoA

ADP+PI ATPCx IICoQ

Cx IIICyto C

Cx IVCx V

LRRK2

Tau

PINK1p64

PINK1p60

CCCP

MG132

LRRK2-IN-1

Caspase-9, Inhibitor I

Caspase-3 Inhibitor V

PINK1MTS

TOM

TOM

ParkinParkin

Accumulation of misfolded pPINK1

DepolarizedMitochondrion

Release ofIMS Proteins

PINK1p64

Abnormal Phosphorylation

Ca2+ Disfunction

DJ1

Apaf1ATP

Cyto C

DOPAMINE

ex2 ex3 ex4 ex5 ex6

NNN +

-20y -10y 0 10y 20y

UbUb

Ub

UbUb

Ub

UbUb

UbUb

UbUb

UbUb

UbUb

UbUb

UbUb

UbUb

UbUb

UbUb

Ub

Ub

UbUb

UbUbUb

HO

NH2

COOH

HO

HO NH2

COOH

L-Dopa

HO

HO NH2

Dopamine

Dopamine o-quinone

Oxidation (H+ + 2e-)

O

O NH2

HO

OH

O

HO

OH

O

OH

HO

O

O

OH

CH3

SuccinateFumerate

VMAT

D2R

DAT

DA

3-MT

HVA

DMHPA

MAO

COMT

ALDH

VSSC CaV 1.2/1.3/2.1/2.2

GIRK

VGCC

Ca2+

Na+ Ca2+

K+

MFNVDACFIS1Ub

UbUb

Ub

TOM

TOM

TIM

TIM

TIMMPP PARL

PINK1p53

MG132SensitiveProtease

TIM

Cyto C

BaxBcl2

p53

Caspases 3,6,7

Caspase 9 p50 p65

IκB

p50 p65

ASC NLRP3

Pro-IL-18

Pro-Caspase-1

JNK/c-Jun kinase

AIF

TRX

TXNIP

Caspase-1

Pro-IL-1β

IL-18IL-1β

?

PINK1p64

DNA Strand Breaks

ATP Depletion

Astrocyte &microglia IL-1

Pro-Inflammatory signals

Quinacrine HCl

Mitophagy

I. II. III. IV. V.

Lewy Bodies

Lewy Neurites

PINK1

MTS TM

A68P A168P H271Q

I II III IV V VI VII VIII IX X XISubstrate Recognition

CN

G309D

L347P

E417G

W437X

LRRK2

ANK LRR

R1441CR1441GR1441H R1628P

ROC COR Kinase WD40

CN

12012T 12020T

Y1699C G2019S G2385R

Parkin

Ubiquitin

V15MR33Q

P37LR42P

A46PAB2E

A92V

KN61N

S167N

M192LM192V

K211RK211N

C212Y

T240RT240N

C253YR256CR271S

R275W

A339ST351P

R366WV380L

D394N

T415N

G430D

C431FP437L

D280NG284R

C289G

Q100H

RING1 IBR RING2

CN

D1R D2R D3R D4R D5R

Platforms & TechnologiesAntibodies & ImmunoassaysEMD Millipore offers an extensive portfolio of antibodies and immunoassays for Alzheimer’s and Parkinson’s disease research. With the expertise of Upstate® and Chemicon®, and backed by excellent service and support, EMD Millipore provides validated and published products in major research areas. Our diverse portfolio includes pathway, cell type, and state-specific antibodies, neurodegenerative markers, stains and assays.

Small Molecules & Cell-based AssaysUse the power of chemical genetics to elucidate mechanisms of neurodegeneration using our wide range of quality, highly published inhibitor and activator small molecules developed by Calbiochem®. Together with our significant portfolio of cell-based activity assays and reporter systems, they facilitate direct and indirect analyses of cell health, migration and neurite extension.

Cells & Cell CultureEMD Millipore’s innovative cell culture solutions help optimize cell growth and maintenance. We offer an extensive range of human and rodent stem cells, primary cells and media designed for most types of stem cells, including embryonic, mesenchymal, and neural stem cells. These optimized media include serum-free, feeder-free formulations, and are supported by our feeder cells, supplements, reagents, sterile filtration and cultureware.

Flow Cytometry Assays & Benchtop SystemsFlow cytometry is an essential tool for in-depth cell analysis, with the capacity to simultaneously measure multiple parameters on individual cells and now high-speed imaging analysis. Our Muse™, guava® easyCyte™ and Amnis® platforms offer a continuum of precision benchtop flow cytometry, from easy, low-cost instruments to high resolution spatial imaging systems. EMD Millipore also offers many specialized reagents and kits for neural stem cell development, cell signaling, oxidative stress and apoptosis analyses in neurodegenerative disease.

MILLIPLEX® map Multiplex AssaysMILLIPLEX® map bead-based neuroscience panels, using trusted Luminex® xMAP® technology, enable multivariate analysis of complex physiologies underlying neuropathology. EMD Millipore offers the widest range of neuroscience related biomarkers. Our assays measure multiple neurodegenerative disease biomarkers simultaneously using small sample volumes.

Your neural connection for antibodies, proteins, kits and assays:www.millipore.com/neuroscience

EMD Millipore and the M mark, Muse, and guava easyCyte are trademarks and MILLIPLEX, Calbiochem, Upstate, and Chemicon are registered trademarks of Merck KGaA, Darmstadt, Germany. Luminex xMAP is a registered trademark of Luminex Corporation.Amnis is a registered trademark of Amnis Corporation. Lit. No. PS4465EN00 09/2012 Job No. LS-SBU-12-06497 Printed in the U.S.A. ©2012 EMD Millipore Corporation, Billerica, MA, USA.

www.emdmillipore.comEMD Millipore is a division of Merck KGaA, Darmstadt, Germany