Apoptosis

Cell Death
The body is very good at maintaining a constant number of cells. So there
has to exist mechanisms for ensuring other cells in the body are removed,
when appropriate.
Mainly Two forms
Apoptosis - suicide - programmed cell death
Necrosis - killing - decay and destruction
Cell Death

1. Apoptosis--Vital process that helps eliminate unwanted

cells--an internally programmed series of events
effected by dedicated gene products also termed as type
I cell death.
Cell death by suicide
-Internal signals (Physical/Chemical insults)
-External signals called Death Factors
Apoptosis Morphology:
Characteristic changes in the nuclear morphology
Overall cell shrinkage
Blebbing of the plasma membrane
Chromatin condensation and precise fragmentation
Formation of apoptotic bodies that contain nuclear
or cytoplasmic material
2. Necrosis--(irreversible injury) changes produced
by enzymatic digestion of dead cellular elements.
Type II cell death.

Cell death by injury

-Mechanical damage
-Exposure to toxic chemicals
 Morphology of necrosis :

1. Cellular swelling or rupture

2. Cell membrane leaky
3. Denaturation and coagulation of
cytoplasmic proteins
4. Breakdown of cell organelles
5. Breakdown of nuclear DNA
6. Cell spills out their contents into
surrounding tissues and cause inflammation
Programmed Cell
Death/Apoptosis
SEEN IN THE FOLLOWING CONDITIONS:
A. Physiologic

1. During development, for removal of excess cells

during embryogenesis

2. To maintain cell population in tissues with high

turnover of cells, such as skin, bowels.

3. To eliminate autoreactive T-cells in developing

thymus.
SEEN IN THE FOLLOWING CONDITIONS:
B. Pathologic

1. To remove damaged cells by virus

2. To eliminate cells after DNA damage by radiation,

cytotoxic agents etc.

3. Cell death in tumors

Apoptosis: liver cells are dying individually from injury by
viral hepatitis.
Molecular Mechanism of Apoptosis

• Proteins which degrade other proteins , employed by

apoptosis - caspases
• The CASP protein is a member of the cysteine-
aspartic acid proteases family. Sequential activation of
caspases plays a central role in the execution-phase
of cell apoptosis.
• Caspase are central to both apoptopoic
pathways,act like molecular scissors to cleave
intracelular protein at aspartate residues.
• Caspases are synthesized as inactive enzymes called pro-caspases
that need to cleave at aspatate residues in order to be activated.
• Caspase cleaves at aspartate residues and the pro-caspases
themselves activated by clevage at aspartate residues.
Factors influencing cell suicide

Withdrawal of positive signals

examples :
growth factors for neurons
Interleukin-2 (IL-2)

Receipt of negative signals

examples :
increased levels of oxidants within the cell
damage to DNA by oxidants
death activators :
• Tumor necrosis factor alpha (TNF-)
• Lymphotoxin (TNF-β)
• Fas ligand (FasL)
The Extrinsic Pathway

• A death ligand/factor such as Fas ligand or TNF is received by

transmembrane death receptor-Fas receptor and TNF
receptor
• Receptor undergoes conformational change and
oligomerizes
• Conformational changes exposes death domains (squares)
located on receptors cytoplasmic tails
• Intracellular adapter proteins bind with the death domain
1. FADD (Fas associated death domain protein)
2. TRADD (TNF receptor associated death domain protein)
• the adaptor transduce the signal from receptor to
several molecules of pro-caspases 8 via death
effector domains DED (triangle)
• Molecules of pro-caspases are now in close
proximity and become activated by self cleavage at
aspartate residues.
• Together the Death ligands receptors,adaptors and
the initiator caspase 8 are called DEATH INDUCING
SIGNAL COMPLEX (DISC)
• Caspases 8 activates other caspases 3,6, and 7 and cleave the specific
proteins and result apoptosis
• Regulation
C-FLIP can bind to adaptor via DED and inhibit caspase 8 recruitment
and activation
Intrinsic Pathway

• Doesn’t depend on external stimuli or death factor

• Inside stimuli-DNA damage or Oxidative stress
• Involve mainly bcl2 family proteins
• All apoptopic proteins are present in between the intermembrane
space of two mitochondrial membrane
Bcl2 family proteins

• 20 members of bcl2 family proteins

• All members contains at least one BH domain
• They act at the outer mitochondrial membrane
• Two groups in Bcl2 family having opposite functions
1. Anti-apoptopic group-inhibit apoptosis
2. Pro-apoptopic group-promotes apoptosis
• Within pro-apoptopic group a subset include BH3
only members those share only BH3 domains
• Their function are
1. either inducing the activity of pro-apoptomic
members
2. Or inhibiting the activity of anti-apoptopic
members
Mechanism of intrinsic pathway

• Apoptopic signal due to Inside stimuli-DNA damage or

Oxidative stress
• The pro-apoptotic bcl-2 proteins are often found in the
cytosol where they act as sensors of cellular damage or
stress.
• Following cellular stress they relocate to the surface of the
mitochondria where the anti-apoptotic proteins are located.
This interaction between pro- and anti-apoptotic proteins
disrupts the normal function of the anti-apoptotic bcl-2
proteins and can lead to the formation of pores in the
mitochondria and the release of cytochrome C and other
pro-apoptotic molecules from the intermembrane space
•
• BAX or BAK undergoes conformational changes
• Oligomerization of 6-8 molecules of BAX /BAK induced
by BH3 family proteins Bid and Bim
• Cytochrome C of e transport chain are combined with
pro-caspase 9 and assembled into a compartment
called APOPTOSOME(Wheel like complex ) along with
Apaf-1(protein cofactor for activation of pro-caspase 9
)
• Caspase 9 activated
• Caspases 9 activates other caspases 3,6, and 7 and cleave the specific
proteins and result apoptosis
• Regulation
Bcl2 anti apoptopic protein,inhibit the association of pro-caspase 9
with Cyt c and Apaf-1 in cytosol
Apoptosis regulating genes
 Apoptosis: Pathways

“Extrinsic Pathway”

Death Death Initiator

Ligands Receptors Caspase 8

Effector
“Intrinsic Pathway”
Caspase 3 PCD

DNA Initiator
Mitochondria/
damage Caspase 9
Cytochrome C
& p53
Apoptosis summary
Necrosis vs. Apoptosis
NECROSIS Vs APOPTOSIS

Wilde, 1999