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Respiration in Yeast and Mammalian Cells http://www.etsu.edu/biology/symbiosis/index Respiration in Yeast and Mammalian Cells

Respiration in Yeast and Mammalian Cells Respiration in Yeast and Mammalian Cells

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Page 1: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Respiration in Yeast and Mammalian Cells

http://www.etsu.edu/biology/symbiosis/index.htm

Respiration in Yeast and Mammalian Cells

Page 2: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

C6H12O6 + 6O2 +6H2O 6CO2 +12H2O

36 ATP/glucose molecule

Respiration: The cellular process by which glucose is oxidized to carbon dioxide and water. An inorganic molecule, often oxygen, serves as the final electron acceptor.The formation of ATP from this process can subsequently be used as the cell’s main source of energy

3 Step Process:1-Glycolysis2-TCA cycle3-Electron transport chain

Glucose

Page 3: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://www.biologycorner.com/resources/resp_summary.jpg

Page 4: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Glycolysis

Page 5: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://fig.cox.miami.edu/~cmallery/150/makeatp/sumgly.jpg

Page 6: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://www.langara.bc.ca/biology/mario/Biol1116notes/biol1116chap6.html

Page 7: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Fermentation:

Page 8: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://www.emc.maricopa.edu/faculty/farabee/BIOBK/BioBookGlyc.html

Page 9: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://www.langara.bc.ca/biology/mario/Biol1116notes/biol1116chap6.html

Page 10: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells
Page 11: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells
Page 12: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://www.emc.maricopa.edu/faculty/farabee/BIOBK/BioBookGlyc.html

Page 13: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells
Page 14: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

http://mgl.scripps.edu/people/goodsell/pdb/pdb72/pdb72_1.html

F0F1 ATP synthase:

Matrix

Inner membrane

Inner membrane space

ADP + Pi ATP

H+ H+H+

Page 15: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Use mitochondria isolated from rat liver cells

Day 2:

Day 1:

Use cultures of Sacchromyces cerevisiae

Page 16: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Glucose

Pyruvate

Acetyl CoA

Acetaldehyde Ethanol

Electron Transport

Glucose and Ethanol Utilization in Yeast

NEM

Azide

Page 17: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Experiment 1: Kinetics of respiration in fresh yeast

Experiment 2: Kinetics of respiration in starved yeast

Experiment 3: Effects of NaN3 on respiration in starved yeast

Experiment 4: Effects of NEM on respiration in starved yeast

Day 1

Page 18: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

•Dissolved oxygen combines with platinum electrons + protons to form water•Electrons move away from the silver electrode; positively charged silverions combine with chloride ions to form silver chloride•The remaining potassium ions combines with the hydroxyl ions from water

The more oxygen there is, the faster the electrons move away from the silver electrode and the more current that is produced

Its very important to keep the stirrer moving!

Teflon membrane

Sample chamber

KClElectrodePlatinum (-)

Silver (+)

The Clark Electrode:

Page 19: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

P/O ratio=ATP synthesized/O2 utilized

AB

C

D

E

F

+m

ed

ium

+m

itos

+substrate

+ADP

Fraction of O2 used

CaCl2

0.4

0.8

1

time in minutes1 2 3 4

>3 indicates tightly coupledmitochondria

Day 2:

You will be testing various combinations of substrates, stimulators, inhibitors, and uncouplers on the rate of respiration

Page 20: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Antimycin A

Glutamate,malate-complex ISuccinate-complex IIAscorbate + TMPD-complex IV

Azide

OligomycinRotenone

http://www.cgm.cnrs-gif.fr/podospora/images/chaine_respi400.jpg

Page 21: Respiration in Yeast and Mammalian Cells  Respiration in Yeast and Mammalian Cells

Many of the chemicals you will be using are toxic

Be Careful!!!!