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INTRINSIC APOPTOSIS PATHWAY Marieta Garib Aisha Green Linda Miranda

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INTRINSIC APOPTOSIS PATHWAY Marieta Garib Aisha Green Linda Miranda. WHAT IS INTRINSIC APOPTOSIS AND WHY DO WE CARE?. Programmed cell-death involving permeability of mitchondria . Involves Caspase-9 As opposed to extrinsic. Tumor necrosis factor Caspase-8 No mitochondria. - PowerPoint PPT Presentation
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INTRINSIC APOPTOSIS PATHWAY Marieta Garib Aisha Green Linda Miranda
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Page 1: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

INTRINSIC APOPTOSIS PATHWAY

Marieta GaribAisha Green

Linda Miranda

Page 2: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

WHAT IS INTRINSIC APOPTOSIS AND WHY DO WE CARE?

•Programmed cell-death involving permeability of mitchondria.Involves Caspase-9•As opposed to extrinsic.Tumor necrosis factorCaspase-8No mitochondria

Page 3: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

WHAT IS INTRINSIC APOPTOSIS AND WHY DO WE CARE?

•Intrinsic pathway induced by chemotherapeutic agents.

•Activation or downregulation of apoptosis influence cancer cell viability.

Page 4: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

http://www.qiagen.com/GeneGlobe/Pathways/Mitochondrial%20Apoptosis.jpg

Page 5: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda
Page 6: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda
Page 7: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

•Caspase 3 Activation = 377.940 seconds•1 Molecule = 477.740 seconds

Page 8: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

•Activation = 394.350 seconds•.99 Caspase-3 = 477.740 seconds

Page 9: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

•Activation = 306.390 seconds•.98 molecule Caspase-3 = 484.900

Page 10: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda
Page 11: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

•Caspase-3 is activated at 250.250 sec.•1 molecule 499.490 sec.

Page 12: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

200.000 250.000 300.000 350.000 400.000 450.000 500.000 550.0000

0.2

0.4

0.6

0.8

1

1.2

Rate of Activation of Caspase 3

10,000 nM dATP

Time (seconds)

Casp

ase

3 M

olec

ules

•Activation = 239.044 secs•1 molecule = 490.759 secs

Page 13: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

•Activation = 229.350 secs•1 molecule = 497.330 secs

Page 14: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

150.000 200.000 250.000 300.000 350.000 400.000 450.000 500.0000.000

0.200

0.400

0.600

0.800

1.000

1.200

Rate of Activation of Caspase 3

20,000 nM dATP

Time (seconds)

Casp

ase

3 M

olec

ules

•Active holoenzyme at 177.137 seconds.•1 molecule of Caspase-3 at 415.351 seconds

Page 15: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda
Page 16: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

Problems and Issues

•choice between intrinsic and extrinsic pathways •finding a target molecule to test

•complexity of the chemical reactions and large number of molecules observed

Page 17: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

Problems and Issues

•difficulty with ODE simulation •long running time of the program due to the complexity of the reactions and the large number of molecules examined

•long t_end=>500 to produce results •very long .gdat files

Page 18: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

CONCLUSIONS

•Varying the concentrations of ATP affects the formation of the holoenzyme

Higher = faster activation of Caspase-3Lower = longer activation time

Page 19: INTRINSIC APOPTOSIS PATHWAY Marieta  Garib Aisha Green Linda Miranda

FUTURE EXPERIMENTS

•Vary concentrations of Apaf-1 and Cytochrome C

•Will they affect holoenzyme formation?


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