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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: The cellular process by which glucose is oxidized to carbon dioxide and

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Respiration in Yeast and Mammalian Cells

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

  2. 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 C6H12O6 + 6O2 +6H2O 6CO2 +12H2O 36 ATP/glucose molecule 3 Step Process: 1-Glycolysis 2-TCA cycle 3-Electron transport chain Glucose

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

  4. Glycolysis

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

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

  7. Fermentation:

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

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

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

  11. H+ H+ F0F1 ATP synthase: H+ Inner membrane space Inner membrane Matrix ADP + Pi ATP http://mgl.scripps.edu/people/goodsell/pdb/pdb72/pdb72_1.html

  12. Day 1: Use cultures of Sacchromyces cerevisiae Day 2: Use mitochondria isolated from rat liver cells

  13. Glucose and Ethanol Utilization in Yeast Glucose NEM Pyruvate Acetaldehyde Ethanol Acetyl CoA Azide Electron Transport

  14. Day 1 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

  15. The Clark Electrode: Sample chamber Teflon membrane KCl Electrode Platinum (-) Silver (+) •Dissolved oxygen combines with platinum electrons + protons to form water •Electrons move away from the silver electrode; positively charged silver ions 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!

  16. Day 2: P/O ratio=ATP synthesized/O2 utilized +medium +mitos 1 +substrate A +ADP B C 0.8 D Fraction of O2 used >3 indicates tightly coupled mitochondria E CaCl2 0.4 F 4 2 1 3 time in minutes You will be testing various combinations of substrates, stimulators, inhibitors, and uncouplers on the rate of respiration

  17. Glutamate,malate-complex I Succinate-complex II Ascorbate + TMPD-complex IV Antimycin A Azide Rotenone Oligomycin http://www.cgm.cnrs-gif.fr/podospora/images/chaine_respi400.jpg

  18. Many of the chemicals you will be using are toxic Be Careful!!!!

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