Cellular Respiration Harvesting Chemical Energy

1. Cellular RespirationHarvesting Chemical Energy ATP

2. What’s thepoint? The pointis to makeATP! ATP

3. Harvesting stored energy • Energy is stored in organic molecules • carbohydrates, fats, proteins • Heterotrophs eat these organic molecules  food • digest organic molecules to get… • raw materials for synthesis • fuels for energy • controlled release of energy • “burning” fuels in a series of step-by-step enzyme-controlled reactions

4. glucose + oxygen  energy + water + carbon dioxide respiration ATP + 6H2O + 6CO2 + heat  C6H12O6 + 6O2 COMBUSTION = making a lot of heat energy by burning fuels in one step ATP glucose O2 O2 fuel(carbohydrates) Harvesting stored energy • Glucose is the model • catabolism of glucose to produce ATP RESPIRATION = making ATP (& some heat)by burning fuels in many small steps ATP enzymes CO2 + H2O + heat CO2 + H2O + ATP (+ heat)

5. + + oxidation reduction e- How do we harvest energy from fuels? • Digest large molecules into smaller ones • break bonds & move electrons from one molecule to another • as electrons move they “carry energy” with them • that energy is stored in another bond, released as heat or harvested to make ATP loses e- gains e- oxidized reduced + – e- e- redox

6. e p loses e- gains e- oxidized reduced + – + + H oxidation reduction H  C6H12O6 + 6O2 6CO2 + 6H2O + ATP H How do we move electrons in biology? • Moving electrons in living systems • electrons cannot move alone in cells • electrons move as part of H atom • move H = move electrons oxidation reduction e-

7. oxidation  C6H12O6 + 6O2 6CO2 + 6H2O + ATP reduction Coupling oxidation & reduction • REDOX reactions in respiration • release energy as breakdown organic molecules • break C-C bonds • strip off electrons from C-H bonds by removing H atoms • C6H12O6CO2 =thefuel has been oxidized • electrons attracted to more electronegative atoms • in biology, the most electronegative atom? • O2H2O =oxygen has been reduced • couple REDOX reactions & use the released energy to synthesize ATP O2

8. Oxidation adding O removing H loss of electrons releases energy exergonic Reduction removing O adding H gain of electrons stores energy endergonic oxidation  C6H12O6 + 6O2 6CO2 + 6H2O + ATP reduction Oxidation & reduction

9. like $$in the bank O– O– O– O– P P P P –O –O –O –O O– O– O– O– O O O O NAD+ nicotinamide Vitamin B3 niacin O O H H C C NH2 C C NH2 How efficient! Build once,use many ways N+ N+ reduction + H oxidation phosphates adenine ribose sugar Moving electrons in respiration • Electron carriers move electrons by shuttling H atoms around • NAD+NADH (reduced) • FAD+2FADH2 (reduced) reducing power! NADH H carries electrons as a reduced molecule

10.  C6H12O6 + 6O2 ATP + 6H2O + 6CO2 Overview of cellular respiration • 4 metabolic stages • Anaerobic respiration 1. Glycolysis • respiration without O2 • in cytosol • Aerobic respiration • respiration using O2 • in mitochondria 2. Pyruvate oxidation 3. Krebs cycle 4. Electron transport chain (+ heat)

11. What’s thepoint? The pointis to makeATP! ATP

12. H+ H+ H+ H+ H+ H+ H+ H+ + P H+ And how do we do that? • ATP synthase enzyme • H+ flows through it • conformational changes • bondPitoADP to make ATP • set up a H+ gradient • allow the H+ to flow down concentration gradient through ATP synthase • ADP + Pi ATP ADP ATP But… How is the proton (H+) gradient formed?

13. H+ H+ H+ H+ H+ H+ H+ H+ + P ADP Got to wait untilthe sequel!Got the Energy?Ask Questions! ATP H+

14. Cellular RespirationStage 2 & 3: Oxidation of Pyruvate Krebs Cycle

15. ATP ATP 2 4 2 2 4 NAD+ ADP ADP 2Pi 2 2Pi 2H glucose C-C-C-C-C-C Overview 10 reactions • convert glucose (6C)to 2 pyruvate (3C) • produces:4 ATP & 2 NADH • consumes:2 ATP • net:2 ATP & 2 NADH fructose-1,6bP P-C-C-C-C-C-C-P DHAP P-C-C-C G3P C-C-C-P pyruvate C-C-C

16. glucose      pyruvate 6C 3C 2x pyruvate       CO2 Glycolysis is only the start • Glycolysis • Pyruvate has more energy to yield • 3 more C to strip off (to oxidize) • if O2 is available, pyruvate enters mitochondria • enzymes of Krebs cycle complete the full oxidation of sugar to CO2 3C 1C

17. Cellular respiration

18. outer membrane intermembrane space inner membrane cristae matrix mitochondrialDNA Mitochondria — Structure • Double membrane energy harvesting organelle • smooth outer membrane • highly folded inner membrane • cristae • intermembrane space • fluid-filled space between membranes • matrix • inner fluid-filled space • DNA, ribosomes • enzymes • free in matrix & membrane-bound What cells would have a lot of mitochondria?

19. Oooooh!Form fits function! Mitochondria – Function Dividing mitochondria Who else divides like that? Membrane-bound proteins Enzymes & permeases bacteria! Advantage of highly folded inner membrane? More surface area for membrane-bound enzymes & permeases What does this tell us about the evolution of eukaryotes? Endosymbiosis!

20. [ ] 2x pyruvate  acetyl CoA + CO2 NAD Oxidation of pyruvate • Pyruvate enters mitochondrial matrix • 3 step oxidation process • releases 2 CO2(count the carbons!) • reduces 2NAD  2 NADH (moves e-) • produces 2acetyl CoA • Acetyl CoA enters Krebs cycle 1C 3C 2C Wheredoes theCO2 go? Exhale!

21. NAD+ 2 x [ ] Pyruvate oxidized to Acetyl CoA reduction Acetyl CoA Coenzyme A CO2 Pyruvate C-C C-C-C oxidation Yield = 2C sugar + NADH + CO2

22. 1937 | 1953 Krebs cycle • aka Citric Acid Cycle • in mitochondrial matrix • 8 step pathway • each catalyzed by specific enzyme • step-wise catabolism of 6C citrate molecule • Evolved later than glycolysis • does that make evolutionary sense? • bacteria 3.5 billion years ago (glycolysis) • free O22.7 billion years ago (photosynthesis) • eukaryotes 1.5 billion years ago (aerobic respiration = organelles  mitochondria) Hans Krebs 1900-1981

23. 2C 6C 5C 4C 3C 4C 4C 4C 4C 6C CO2 CO2 Count the carbons! pyruvate acetyl CoA citrate oxidationof sugars This happens twice for each glucose molecule x2

24. 2C 6C 5C 4C 3C 4C 6C 4C 4C 4C NADH ATP CO2 CO2 CO2 NADH NADH FADH2 NADH Count the electron carriers! pyruvate acetyl CoA citrate reductionof electroncarriers This happens twice for each glucose molecule x2

25. Whassup? So we fully oxidized glucose C6H12O6  CO2 & ended up with 4 ATP! What’s the point?

26. H+ H+ H+ H+ H+ H+ H+ H+ H+ Electron Carriers = Hydrogen Carriers • Krebs cycle produces large quantities of electron carriers • NADH • FADH2 • go to Electron Transport Chain! ADP+ Pi ATP What’s so important about electron carriers?

27. 4 NAD+1 FAD 4 NADH+1FADH2 2x 1C 3x 1 ADP 1 ATP Energy accounting of Krebs cycle Net gain = 2 ATP = 8 NADH + 2 FADH2 pyruvate          CO2 3C ATP

28. Value of Krebs cycle? • If the yield is only 2 ATP then how was the Krebs cycle an adaptation? • value of NADH & FADH2 • electron carriers & H carriers • reduced molecules move electrons • reduced molecules move H+ ions • to be used in the Electron Transport Chain like $$in the bank

29. What’s thepoint? The pointis to makeATP! ATP

30. H+ H+ H+ H+ H+ H+ H+ H+ + P H+ And how do we do that? • ATP synthase • set up a H+ gradient • allow H+ to flow through ATP synthase • powers bonding of Pi to ADP ADP + PiATP ADP ATP But… Have we done that yet?

31. NO!The final chapter to my story is next! Any Questions?

32. Cellular RespirationStage 4: Electron Transport Chain

33. Cellular respiration

34. What’s thepoint? The pointis to makeATP! ATP

35. ATP accounting so far… • Glycolysis 2ATP • Kreb’s cycle 2ATP • Life takes a lot of energy to run, need to extract more energy than 4 ATP! There’s got to be a better way! I need a lotmore ATP! A working muscle recycles over 10 million ATPs per second

36. O2 There is a better way! • Electron Transport Chain • series of proteins built into inner mitochondrial membrane • along cristae • transport proteins& enzymes • transport of electrons down ETC linked to pumping of H+ to create H+ gradient • yields ~36 ATP from 1 glucose! • only in presence of O2 (aerobic respiration) Thatsounds morelike it!

37. Mitochondria • Double membrane • outer membrane • inner membrane • highly folded cristae • enzymes & transport proteins • intermembrane space • fluid-filled space between membranes Oooooh!Form fits function!

38. Innermitochondrialmembrane Electron Transport Chain Intermembrane space C Q cytochromebc complex cytochrome coxidase complex NADH dehydrogenase Mitochondrial matrix

39. Remember the Electron Carriers? glucose Krebs cycle Glycolysis G3P 8 NADH 2 FADH2 2 NADH Time tobreak openthe piggybank!

40. e p 1 2 Electron Transport Chain Building proton gradient! NADH  NAD+ + H intermembranespace H+ H+ H+ innermitochondrialmembrane H  e- + H+ C e– Q e– H e– FADH2 FAD H NADH 2H+ + O2 H2O NAD+ cytochromebc complex cytochrome coxidase complex NADH dehydrogenase mitochondrialmatrix What powers the proton (H+) pumps?…

41. H+ H+ H+ H+ H+ H+ H+ H+ H+ H+ H+ H+ C e– Q e– 1 2 e– FADH2 FAD NADH 2H+ + O2 H2O NAD+ cytochromebc complex NADH dehydrogenase cytochrome coxidase complex Stripping H from Electron Carriers • Electron carriers pass electrons & H+ to ETC • H cleaved off NADH & FADH2 • electrons stripped from H atoms  H+ (protons) • electrons passed from one electron carrier to next in mitochondrial membrane (ETC) • flowing electrons = energy to do work • transport proteins in membrane pump H+ (protons) across inner membrane to intermembrane space H+ H+ H+ TA-DA!! Moving electronsdo the work! ADP+ Pi ATP

42. H2O O2 But what “pulls” the electrons down the ETC? electronsflow downhill to O2 oxidative phosphorylation

43. Electrons flow downhill • Electrons move in steps from carrier to carrier downhill to oxygen • each carrier more electronegative • controlled oxidation • controlled release of energy make ATPinstead offire!

44. H+ H+ H+ H+ H+ H+ H+ H+ ADP + Pi H+ “proton-motive” force We did it! • Set up a H+ gradient • Allow the protonsto flow through ATP synthase • Synthesizes ATP ADP + PiATP ATP Are wethere yet?

45. Chemiosmosis • The diffusion of ions across a membrane • build up of proton gradient just so H+ could flow through ATP synthase enzyme to build ATP Chemiosmosis links the Electron Transport Chain to ATP synthesis So that’sthe point!

46. 1961 | 1978 Peter Mitchell • Proposed chemiosmotic hypothesis • revolutionary idea at the time proton motive force 1920-1992

47. Intermembrane space Pyruvate from cytoplasm Inner mitochondrial membrane H+ H+ Electron transport system C Q NADH e- H+ 2. Electrons provide energy to pump protons across the membrane. 1. Electrons are harvested and carried to the transport system. e- Acetyl-CoA NADH e- H2O e- Krebs cycle 3. Oxygen joins with protons to form water. 1 FADH2 O2 2 O2 + 2H+ H+ CO2 ATP H+ ATP ATP 4. Protons diffuse back indown their concentrationgradient, driving the synthesis of ATP. ATP synthase Mitochondrial matrix

48. ~40 ATP Cellular respiration + + 2 ATP 2 ATP ~36 ATP

49.  C6H12O6 + 6CO2 + 6H2O + ~40 ATP 6O2 Summary of cellular respiration • Where did the glucose come from? • Where did the O2 come from? • Where did the CO2 come from? • Where did the CO2 go? • Where did the H2O come from? • Where did the ATP come from? • What else is produced that is not listed in this equation? • Why do we breathe?

50. H+ H+ H+ C e– Q e– 1 2 e– FADH2 FAD NADH 2H+ + O2 H2O NAD+ cytochromebc complex NADH dehydrogenase cytochrome coxidase complex Taking it beyond… • What is the final electron acceptor in Electron Transport Chain? O2 • So what happens if O2 unavailable? • ETC backs up • nothing to pull electrons down chain • NADH & FADH2 can’t unload H • ATP production ceases • cells run out of energy • and you die!