CASPASE ACTIVATION

& APOPTOSIS

CASPASE CLEAVAGE & ACTIVATION

Pro-Domain

Large Subunit (p20)

Pro-Domain

chain

Asp-x
Proteolytic cleavage
chain

Pro-Domain

subdivided into three functional groups, apoptotic initiator caspases (Caspase-2, -8, -9, -10), apoptotic eector caspases (Caspase-3, -6, -7), and

chain

TRAIL

chain

Apoptotic caspases are

re activated upon the receipt of either an extrinsic or an intrinsic death signal. The extrinsic

Fas Ligand

TRAIL R1
TRAIL R2

pathway (green arrows)

ws) is initiated by ligand binding to cell surface death receptors (TNF RI, Fas/CD95, DR3,
TRAIL R1/DR4, TRAIL R2/DR5) followed by receptor oligomerization and cleavage of Pro-caspase-8 and -10.

FADD

MAMMALIAN CASPASE DOMAINS & CLEAVAGE SITES

APOPTOTIC CASPASES

FADD

Activation of Caspase-8
-8 and Caspase-10 results in the cleavage of BID and downstream eector

INITIATOR CASPASES

Pro-caspase-8, -10

TWEAK

Pro-caspase-8, -10

caspases. The intrinsicc pathway of caspase activation (purple arrows) is initiated by events such as

Caspase-2

Fas/CD95

152

316 331

FADD

216

374 385

TNF-

brane that result in the

e release of

Pro-caspase-8, -10

pro-apoptotic proteinss including

138

315 331

Inducing Factor (AIF), and

Endonuclease G.

Extrinsic Pathway

219

415

28

175

TRADD
FADD

ARC

Pro-caspase-8, -10

DD
DED

RIP1

Caspase-10

416

CARD

Caspase-9

DED

521

DED

tBID
Pro-caspase-8, -10

Death Domain (DD)

Mitochondrial membrane
rane permeability is regulated by the

BID

TRADD

TRADD
TRAF-2

479

DED

DR3 (or another TWEAK R)

Caspase-8, -10
RIP1
TRADD
TRAF-2

DED

Caspase-8

Pro-caspase-8, -10
FLIP

TNF RI

FADD

435

CARD

DNA damage, growth factor withdrawal, or loss of contact with the extracellular matrix. These

HTRA2/Omi, Apoptosisis-

chain
Active caspase

containing a variable length pro-domain, followed by a large (20 kDa) and a small (10 kDa) subunit.

Cytochrome c, Smac/Diablo,
Diablo,

chain

Heterotetramer
Formation

Caspases are a family of aspartate-specic, cysteine proteases that serve as the primary mediators of apoptosis. Mammalian caspases can be

events lead to changes

es in the integrity of the mitochondrial mem-

chain

Asp-x

Extrinsic & Intrinsic Pathways of Caspase Activation

caspases involved in inammatory

nammatory cytokine processing (Caspase-1, -4, -5, 11, and -12L/12S). All caspases are synthesized as inactive zymogens

Small Subunit (p10)

BAK

Cytochrome c
SMAC/Diablo
HTRA2/Omi

Death Eector Domain (DED)

Bcl-2 family of proteins.

ns. The balance between pro- and
anti-apoptotic family members determines whether or
not a cell will undergo
o apoptosis. In healthy cells,

tBID

tBID

Bad

14-3-3
P

Bad
BAK

tBID

BAK

IAPs

tBID

EFFECTOR CASPASES

IAPs

Caspase-9

Bcl-2

Bcl-2 and Bcl-xL inhibitt apoptosis by binding to the

BAK

Bcl-xL

APAF-1

Pro-caspase-9
Apoptosome

tBID

protein in these cells. If cytoplasmic levels of free BAD increase, Bcl-2

BAK

and Bcl-xL bind to Bad and release Bax and BAK. Bax and BAK, or processed
forms of these proteins,
ns, can then insert into the mitochondrial membrane,

tBID

Cytochrome c

compromising its integrity.

grity.

Bcl-2

Bcl-xL
Bad
Bax

14-3-3

Caspase-3

Pro-caspase-7
Caspase-7

Target Molecules
(e.g. Actin,
Nuclear lamins,
ICAD, PARP)

protein, FADD. This interaction

teraction is mediated by

cleavage and activation.

on. In

Bcl-2
Bax

tBID

Bcl-2

Pro-caspase-6

tBID

Bad

tBID
Bax

Caspase-6

Cytochrome c

Cytochrome c
AIF

270 290

134

311 331

ARC

373

Endo G

CARD

Caspase-11

Caspase-2

PIDDosome

Caspase-12S

DNA FRAGMENTATION

Caspase-12L

Pro-caspase-2

125

CARD

419

CARD

CASPASE ACTIVATING COMPLEXES

CRADD/RAIDD
DISC

DNA DAMAGE

CARD

x x x
x x

PIDD
DD

p53

p53

p53

activated following an
n

INTERACTING DOMAINS

INHIBITORS OF COMPLEX FORMATION

Fas Ligand
Fas/CD95

Pro-apoptotic: Anti-apoptotic:
Bad
Bax
BID
Noxa

PUMA
PIDD
others

14-3-3
p21
others

FADD
Pro-caspase-8,-10
DED

DD
DD/DED
DED

DD

FLIP

Intrinsic Pathway

associated with

APOPTOSOME

INTERACTING DOMAINS

the release of
Cytochrome c from the
e mitochondria. In the cytoplasm, Cytochrome c interacts with APAF-1, which

418

CARD

Caspase-5

BID

377

CARD

Caspase-4

HSPs

contrast, Pro-caspase-9
-9 is
intrinsic change

93

404

CARD

Caspase-1

DD

death receptors results

ts in their

297 317

INFLAMMATORY CASPASES

Cytochrome c

their shared death eector

ector domains (DED).
Clustering of pro-caspases
pases near the

119

303

Caspase-7

Pro-caspase-8 and Pro-caspase-10

o-caspase-10 are recruited to the death
receptors through their
eir interactions with the adaptor

198

Bad

23

293

Bax
Bcl-xL

Bad

inducing signaling complex

mplex (DISC; Caspase-8 and -10), the apoptosome
following ligand binding
ing and death receptor oligomerization.

tBID

Bax

Initiator caspases are activated in three distinct protein complexes, the death(Caspase-9), and the PIDDosome (Caspase-2). The DISC is formed

Bax

Bcl-xL
BAK

Bad

Pro-caspase-3

Caspase Recruitment
Domain (CARD)

Bcl-xL

pro-apoptotic Bax and

d BAK proteins. Bad is also phosphorylated and sequestered
estered in the cytoplasm by the 14-3-3

179 194

Caspase-6

Bad

23

Bax

Bcl-2

277

Caspase-3

ARC

INHIBITORS OF COMPLEX FORMATION

HSP27

NUCLEUS

HSP70

Cytochrome c

recruits Pro-caspase-9
9 by way of its caspase recruitment domain (CARD) to form the apoptosome.

APAF-1

CARD

Pro-caspase-9

CARD

HSP27

Cytochrome c

Formation of the apoptosome

ptosome leads to the cleavage and activation of Caspase-9. Intrinsic cellular
changes can also lead to the activation of Caspase-2. Following DNA damage, p53 induces the

APAF-1

expression of p53-induced
uced protein with a death domain (PIDD), which associates with the
PIDDOSOME

CRADD/RAIDD adaptor
or protein and Pro-caspase-2 to form the PIDDosome. The association

INTERACTING DOMAINS
PIDD

between CRADD/RAIDD
DD and PIDD is mediated by their shared death domains (DD), while

DD

CARD domains mediate

te the interaction between CRADD/RAIDD and Pro-caspase-2.

CRADD/RAIDD

DD/CARD

Formation of the PIDDosome

Dosome leads to the cleavage of Pro-caspase-2.

Pro-caspase-2

CARD

Autocatalytic cleavage
e of the initiator pro-caspases occurs at aspartic acid residues

INFLAMMASOME

INTERACTING DOMAINS

located after the pro-domain,

domain, and in between the large and the small subunits.
NALP3, NALP1 or IPAF

Pyrin

Upon cleavage, mature

re caspases form a proteolytically active heterotetramer
consisting of two smallll and two large subunits. Once activated, initiator
caspases cleave downstream
nstream eector caspases that promote the ordered
disassembly of the cellll b
by targeting a number
b off criticall cellular
ll l proteins including
l d
structural proteins, DNA repair proteins, and proteins involved in signal transduction pathways.

DOMAIN KEY
Death Domain (DD)

ASC

R&D Systems, Inc. | 1-800-343-7475 | www.RnDSystems.com

Note: This poster conveys a general overview and should be considered neither comprehensive nor denitive. The details of the process are understood
to be subject to interpretation. R&D Systems, Inc. 2010

Pro-caspase-1

Pyrin/CARD

CARD

Death Eector Domain

(DED)
Caspase Recruitment
Domain (CARD)
Pyrin Domain