CH 6: Pathways that Harvest and Store Chemical Energy Mitochondrion Outer membrane Inner membrane Cristae Matrix TEM 44,880 Intermembrane space For HW, read section 6.1 and define the following vocabulary terms: 1. 2. 3. 4. 5. 6. Reduction Oxidation Substrate-level phosphorylation Oxidative phosphorylation Chemiosmosis ATP synthase (2/4) BR: Energy review 1. 2. 3. Describe the difference between endergonic and exergonic reactions. Describe the difference between anabolic and catabolic reactions. What is the energy molecule that cells use to do work? 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism Chemical energy available to do work is termed free energy (G) Five principles governing metabolic pathways: 1. Chemical transformations occur in a series of intermediate reactions – a metabolic pathway or cascade 2. Each reaction is catalyzed by a specific enzyme 3. Most metabolic pathways are similar in all organisms 4. In eukaryotes, many metabolic pathways occur inside specific organelles 5. Each metabolic pathway is controlled by enzymes that can be inhibited or activated 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism Energy coupling: an energy-releasing (exergonic) reaction provides energy for an energy-storing (endergonic) reaction ATP (adenosine triphosphate) ◦ Energy released by exergonic reactions is stored in the bonds of ATP ◦ When ATP is hydrolyzed, free energy is released to drive endergonic reactions ATP H2O ADP Pi freeenergy (Pi refers to an inorganic phosphate molecule) Energy Coupling – energy released from exergonic reactions is used to drive endergonic reactions Formation of ATP requires energy (endergonic) Hydrolysis of ATP releases energy (exergonic) ATP (adenosine triphosphate) Which bond contains more free energy (G), the bond between phosphate groups, or the O—H bond that forms after hydrolysis of ATP? 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism Redox (oxidation–reduction) reactions transfer energy ◦ Reduction is the GAIN of one or more electrons ◦ Oxidation is the LOSS of one or more electrons (“OIL RIG”…oxidation involves loss, reduction involves gain!) Oxygen is one of the strongest oxidizing agents due to its electronegativity! It loves to TAKE electrons from other molecules, thus oxidizing them. 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism ◦ Transfer of hydrogen atoms involve transfer of electrons (H = H+ + e–) ◦ Energy in the reducing agent is transferred to the reduced product When the reducing agent (compound A) gives up electrons, it is oxidized When the oxidizing agent (compound B) takes electrons, it is reduced Oxidation, Reduction, and Energy 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism ◦ Coenzyme NAD+ is a key electron carrier in redox reactions (other examples are FAD & NADP+) Reduction of NAD+ forms NADH Formation of NADH is highly endergonic NAD H 2e NADH ◦ In catabolic processes, oxidation releases energy that is trapped by the reduction of coenzymes such as NADH ◦ Energy for anabolic processes is supplied by ATP NAD+ NADH 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism Oxidative phosphorylation transfers energy from NADH to ATP NADH NAD H 2e energy energy ADP Pi ATP ◦ Achieved via chemiosmosis—the diffusion of H+ (protons) across a membrane to drive the synthesis of ATP H+ diffuse through membrane protein ATP synthase *Based on your vocab and your notes, draw a diagram of what occurs during chemiosmosis How do H+ diffuse through a membrane? Chemiosmosis through ATP Synthase Energy from electron carriers is used to actively transport H+ against its concentration gradient Diffusion of H+ through ATP synthase provides energy for ATP formation (2/4) Exit slip Describe the energy transfer that takes place when ATP is formed versus when it is hydrolyzed. When is energy stored? When is it released? If you can, draw a diagram showing ATP’s role in energy coupling. (1/30) BR: Vocabulary matching the diffusion of protons (H+) across a membrane to drive the synthesis of ATP 2. formation of ATP involving direct transfer of phosphate to ADP 3. the gaining of one or more electrons 4. the loss of one or more electrons 5. energy requiring/absorbing 6. energy-releasing 7. membrane protein through which H+ diffuse to form ATP 8. formation of ATP involving transfer of energy from NADH to ADP 1. (2/8) Bellringer: ATP & respiration What does the word respiration mean? 2. How do photosynthesis and respiration cycle carbon through the biosphere? Refer to the carbon-containing molecules in each process. 3. Copy the diagram of ATP below (just the ATP) and point out where in the molecule the energy is stored. 4. Describe the energy transfer that takes place between the coenzymes ATP & NADH during oxidative phosphorylation (check your notes). Adenosine Triphosphate Adenosine diphosphate Exit review 1. Phosphate P Adenine P P H2O Hydrolysis P P ADP + Pi Ribose ATP Pi Energy BR: Energy review Energy flows through the biosphere ◦ Enters as light and exits as heat Chemicals (matter) are recycled through the biosphere 2. Photosynthesis uses CO2 to build organic molecules like glucose (C6H12O6). Respiration breaks down organic molecules and returns CO2 to the atmosphere. 6.1 ATP, Reduced Coenzymes, and Chemiosmosis Play Important Roles in Biological Energy Metabolism Cellular respiration is a major catabolic pathway in which glucose is oxidized and energy is released: carbohydra te 6O2 6CO2 6H 2O chemical energy Photosynthesis is a major anabolic pathway in which light energy is converted to and stored as chemical energy (glucose): 6CO2 6H 2O light energy 6O2 carbohydra te 6.2 Carbohydrate Catabolism in the Presence of Oxygen Releases a Large Amount of Energy Cellular Respiration – release of energy through the oxidation of glucose (or other organic molecules) ◦ Aerobic respiration occurs in the presence of oxygen I. Glycolysis II. Pyruvate Oxidation III. Citric Acid (Krebs) cycle IV. Electron Transport Chain Cellular Respiration: a series of small steps Why doesn’t the oxidation of glucose occur in one single step? Why doesn’t the oxidation of glucose occur in one single step? ◦ It would be too explosive and not enough energy would be harnessed ◦ There are other oxidizing agents for respiration in the electron transport chain besides oxygen (the oxidizing agent is the recipient of the electrons and therefore is responsible for the oxidation of another molecule) 6.2 Carbohydrate Catabolism in the Presence of Oxygen Releases a Large Amount of Energy Glycolysis – conversion of 6-C glucose into two 3-C pyruvate molecules ◦ Occurs in cytosol ◦ Products: 2 pyruvate, 2 ATP, 2 NADH ATP is produced via substrate-level phosphorylation Glycolysis converts glucose into 2 molecules of pyruvate • Requires an “investment” of 2 ATP at beginning …phosphorylated glucose is more reactive AND cannot exit the cell Glycolysis converts glucose into 2 molecules of pyruvate Glycolysis converts glucose into 2 molecules of pyruvate • Exergonic reactions at the end of glycolysis produce 2 NADH and 4 ATP. ATP are produced via substrate-level phosphorylation. Oxidative phosphorylation via chemiosmosis Substrate level phosphorylation 6.2 Carbohydrate Catabolism in the Presence of Oxygen Releases a Large Amount of Energy Pyruvate Oxidation: links glycolysis & citric acid cycle ◦ Occurs in mitochondrial matrix (2x per glucose) ◦ Products (total): 2 NADH, 2 CO2, & 2 acetyl CoA pyruvate is oxidized to acetate which is then bound to Coenzyme A (CoA), forming acetyl CoA Mitochondrion Outer membrane Intermembrane space Cristae Matrix TEM 44,880 Inner membrane Formation of Acetyl CoA Citric Acid (Krebs) Cycle ◦ Occurs in mitochondrial matrix (2x per glucose) ◦ Products: 4 CO2, 6 NADH, 2 FADH2, & 2 ATP ATP is formed from GTP via substrate-level phosphorylation 2-C acetyl group gets oxidized, then joins the 4-C oxaloacetate to form 6-C citrate (citric acid) GTP can transfer its high-energy phosphate to form ATP 6.2 Carbohydrate Catabolism in the Presence of Oxygen Releases a Large Amount of Energy Electron Transport Chain: NADH & FADH2 are oxidized to NAD+ & FAD, and O2 is reduced to form H2O ◦ Series of redox carrier proteins (the respiratory chain) embedded in the inner membrane of mitochondria Transport the electrons donated by NADH and FADH2 from one carrier to the next Actively transport H+ into intermembrane space, setting up a proton gradient ◦ Proton motive force: H+ diffuse through ATP synthase to synthesize ATP by chemiosmosis 6.2 Carbohydrate Catabolism in the Presence of Oxygen Releases a Large Amount of Energy ◦ Oxygen is the final electron acceptor at the end of the ETC ◦ Products: 6 H2O, 32 ATP ATP is produced via oxidative phosphorylation ◦ Complete chemical equation for aerobic respiration: C6H12O6 + 6O2 6CO2 + 6H2O + Energy (36 ATP) Electron Transport Chain & Chemiosmosis ETC and chemiosmosis animation Intermembrane space Proteins pass electrons from NADH & FADH2. The energy from the electrons is used to pump H+ out of the matrix across the inner membrane of the mitochondrion. O2 is the final electron acceptor at the end of the ETC Diffusion of H+ thru ATP synthase produces ATP via oxidative phosphorylation Aerobic Respiration animation recap Electrons carried via NADH PYRUVATE OXIDATION (2/9) Group BR Aerobic Respiration: C6H12O6 + 6O2 6CO2 + 6H2O + Energy (36 ATP) Cellular respiration involves the relocation of electrons, which releases stored energy in organic molecules like glucose. This energy is used to synthesize ATP. In the process, glucose is oxidized and oxygen is reduced. 1. What molecules does glucose ultimately become once it is fully oxidized during aerobic respiration? 2. What does oxygen become when it is reduced at the end of the ETC? 3. The ATP from aerobic respiration represents ~40% of the energy that was originally stored in glucose. What do you think happens to the other 60%? ◦ Newborns contain a protein called thermogenin that disrupts the H+ gradient in brown fat, preventing ATP formation and increasing the release of heat. 4. Dinitrophenol (DNP) is an “uncoupler,” which means it interferes with the flow of electrons during electron transfer and allows H+ to leak back into the matrix across the inner membrane (see figure below). Fifty years ago, DNP was given as a drug to help patients lose weight. a. Why would taking DNP make someone lose weight? b. Why would taking DNP be dangerous? Intermembrane space Inner membrane Mitochondrial Matrix 6.3 Carbohydrate Catabolism in the Absence of Oxygen Releases a Small Amount of Energy Anaerobic respiration occurs entirely in the cytosol in the absence of O2 I. Glycolysis (2 ATP) II. Fermentation (0 ATP) Fermentation regenerates NAD+ so glycolysis can continue ◦ Lactic Acid Fermentation – NADH reduces pyruvate to lactic acid Performed by animals, plants, microorganisms,… Buildup of lactic acid in muscles inhibits muscle contraction ◦ Alcoholic Fermentation – pyruvate is converted to ethanol and CO2 Performed by yeast Alcoholic and lactic acid fermentation * In both cases NAD+ is regenerated from NADH Stored ATP and creatine phosphate (CP) can provide 10-15 seconds worth of energy to muscles. Anaerobic respiration provides energy for a limited time before lactic acid buildup begins to inhibit muscle contractions. 6.4 Catabolic and Anabolic Pathways Are Integrated Carbon skeletons (molecules with covalently linked carbon atoms) can enter catabolic or anabolic pathways Catabolism ◦ Polysaccharides are hydrolyzed to glucose, which enter glycolysis ◦ Triglycerides (lipids) break down to fatty acids and glycerol. Fatty acids can be converted to acetyl CoA & enter the Krebs cycle ◦ Proteins are hydrolyzed to amino acids that can feed into glycolysis or the Krebs cycle 6.4 Catabolic and Anabolic Pathways Are Integrated Anabolism ◦ Gluconeogenesis—Krebs cycle and glycolysis intermediates can be reduced to form glucose ◦ Acetyl CoA can be used to form fatty acids ◦ Some citric acid intermediates can form nucleic acids (2/10) BR (Quiz tomorrow!) A. B. (Stage) Cytoplasm C. D. (Stage) E. F. (stage) Mitochondrial matrix * Write down any specific questions you have regarding cellular respiration. 1. Identify the stages or molecules in the diagram. 2. Which type of respiration is shown in the diagram, aerobic or anaerobic? G. (stage) CO2 and H2O Inner mitochondrial membrane A. Glucose B. Glycolysis Cytoplasm C. Pyruvate D. Pyruvate oxidation E. Acetyl CoA F. Krebs cycle Mitochondrial matrix G. ETC CO2 and H2O Inner mitochondrial membrane Aerobic Respiration animation recap Electrons carried via NADH PYRUVATE OXIDATION 3. The evolution of photosynthesizing organisms and the development of an O2-rich environment led to a rapid diversification of life. Explain why there is an evolutionary advantage to an organism that requires oxygen to live compared to one that does not require oxygen. 4. Muscle fatigue results from EITHER lactic acid accumulation deactivating muscle contractions OR muscles running out of energy reserves. Identify which you would expect to happen during a marathon versus a sprint and WHY. Effect of O2 Levels on Evolution Label the mitochondrion below. Letter “a” is pointing to the inward fold. a b c d e c. er mbrane d. a. b. e. (2/10) BR: Photosynthesis intro 5. 6. 7. 8. What similarities do mitochondria and chloroplasts share? What are the reactants and products of photosynthesis? Why do plants perform photosynthesis? What types of organisms rely on photosynthesis for survival? 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy Photosynthesis: energy from sunlight is stored as chemical energy in glucose Light ◦ 6CO2 + 6H2O C6H12O6 + 6O2 ◦ 2 pathways I. Light reactions convert light energy into chemical energy (ATP and NADPH) II. Carbon-fixation reactions (Calvin cycle) use the ATP and NADPH to produce glucose from CO2 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy Light is a form of electromagnetic radiation, which travels as a wave but also behaves as particles (photons) ◦ When a molecule absorbs photons, its electrons become energized or “excited” Generally, the excited electron will fall back down and release energy as heat…in isolation, chlorophyll releases energy as heat and light Fluorescence Chloroplasts are green because chlorophyll REFLECTS green wavelengths of light. Other wavelengths of light in the visible spectrum are absorbed (2/11) BR The graph shows the absorption and action spectra of chlorophyll a, the main pigment involved in photosynthesis. An absorption spectrum shows which wavelengths of light are absorbed, and an action spectrum shows the amount of photosynthetic activity at different wavelengths of light. 1. What colors does chlorophyll a absorb? 2. What is the relationship between light absorption and photosynthetic activity? 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy Pigments: molecules that absorb light ◦ Absorption spectrum—shows light energy absorbed at different wavelengths ◦ Action spectrum— shows biological activity of an organism at different wavelengths 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy ◦ Chlorophyll absorbs blue and red light and reflects green ◦ Accessory pigments (e.g., betacarotene) transfer energy to & protect chlorophyll ◦ Pigments are arranged into light-harvesting complexes, or antenna systems ◦ A photosystem consists of antenna systems & a reaction center If chl a is the only pigment that actually passes the light energy to electrons to start photosynthesis, then how is it that there is photosynthesis at 500 nm of light? If the chl a is the only pigment that actually passes the light energy to electrons, then how is it that there is photosynthesis at 500 nm of light? • Accessory pigments absorb light too! Photosystem Based on the diagrams, what purpose does a long hydrocarbon tail serve to a chlorophyll molecule? What other macromolecule have you studied that has a hydrocarbon tail? CHLOROPHYLL Fatty acids in the thylakoid membrane are hydrocarbons! The thylakoid membrane is a phospholipid bilayer just like a cell membrane, so the hydrocarbon tail on chlorophyll anchors it within the bilayer. Phospholipid Chlorophyll Photosystem Photosystems are embedded CHLOROPHYLL within integral membrane proteins and consist of antenna systems surrounding a reaction center Why do the leaves Turn Color in the Fall? What causes the leaves to be green to begin with? Chlorophyll a and b The colors that we see are a reflection of the wavelengths of light that the pigments cannot absorb. Why do leaves turn colors in the fall? Shorter days and cooler nights stop chlorophyll production and drive chlorophyll out of leaves Accessory pigments normally masked by chl are revealed High amount of sugar in leaves leads to production of anthocyanins (red/purple/blue) Why do the leaves fall off of the trees, and die soon after the color change? Without chl, no photosynthesis, and veins carrying water & nutrients to/from leaves are closed off (2/12) BR: Photosynthesis 1. 2. Write the chemical equation for photosynthesis. Then label which reactant is oxidized and which reactant is reduced. Consider the requirements plants need to grow & survive. Where does most of the organic material forming the biomass of a plant come from? (2/12) BR 2. Identify the inner membrane, outer membrane, thylakoid, granum, and stroma. a. b. c. d. e. (2/12) Bellringer a. granum b. thylakoid Copyright Pearson Prentice Hall c. Outer mem. d. Inner mem. e. stroma 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy I. Light reactions – take place in the thylakoid membrane ◦ At photosystems chlorophyll absorbs light and releases an excited electron Photosystem II – electron is passed to an electron acceptor in the ETC Water is split to replace the electron in chlorophyll (photolysis), producing O2 Chemiosmosis produces ATP (photophosphorylation) Photosystem I – electron passes to NADP+ (the final electron acceptor) which is reduced to form NADPH 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy ◦ Noncyclic electron transport uses 2 photosystems Electron from PS II replaces excited electron in PS I Electron transport chains produce ATP & NADPH Noncyclic Electron Flow Noncyclic Electron Transport 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy ◦ Cyclic electron transport uses photosystem I and produces additional ATP for carbon-fixation reactions Excited electron cycles back to chlorophyll Cyclic Electron Flow 6.6 Photosynthetic Organisms Use Chemical Energy to Convert CO2 to Carbohydrates Calvin cycle (carbon-fixation reactions) – takes place in the stroma ◦ 3 steps: 1. CO2 Fixation CO2 is added to RuBP (ribulose 1,5-bisphosphate) in a reaction catalyzed by the enzyme rubisco Produces two 3PG (3-phosphoglycerate) molecules 2. Reduction: 3PG is reduced to form G3P (glyceraldehyde 3-phosphate) 3. Regeneration: RuBP is regenerated 6.6 Photosynthetic Organisms Use Chemical Energy to Convert CO2 to Carbohydrates Rubisco catalyzes a reaction that combines CO2 with RuBP forming 3phosphoglycerate (3PG) RuBP is regenerated from RuMP to complete the cycle 3PG is reduced to glyceraldehyde 3phosphate (G3P) in a 2step reaction About 1/6 of G3P molecules are used to make sugars 6.5 During Photosynthesis, Light Energy Is Converted to Chemical Energy ◦ Extra G3P is exported to the cytosol and converted to hexoses (glucose and fructose) Glucose molecules are linked together to form starch or cellulose The C—H bonds generated by the Calvin cycle provide almost all the energy for life on Earth ◦ Photosynthetic organisms (autotrophs) use most of this energy in cellular respiration to support their own growth and reproduction ◦ Heterotrophs depend on autotrophs for chemical energy (i.e., food) to harvest during cellular respiration Photosynthesis Video Animation Hydrogen bonding to other cellulose molecules can occur at these points The G3P made in the Calvin Cycle is involved in the biosynthesis of other organic molecules 1. 2. 3. Explain the effect of increasing light intensity on the rate of photosynthesis. Explain the effect of increasing temperature on the rate of photosynthesis. Explain WHY the rate of photosynthesis is affected in these ways. (Do the shapes of these graphs look familiar?) Effect of Temperature on Rate of Photosynthesis Rate of Photosynthesis (2/12) Exit Effect of Light Intensity on Rate of Photosynthesis 100 80 60 40 20 0 20 40 60 Temp (C) 80 100 (2/16) BR 1. Get out your notes from CH 6 and write down any concepts you are struggling to understand (oxidation, reduction, energy coupling, substrate-level/oxidative phosphorylation, chemiosmosis, glycolysis, pyruvate oxidation, Krebs cycle, ETC, fermentation, pigments, light reactions, photolysis, photophosphorylation, Calvin cycle, carbon fixation…) Effect of Light Intensity on Rate of Photosynthesis Rate of Photosynthesis Effect of Temperature on Rate of Photosynthesis 100 80 60 40 20 0 20 40 60 Temp (C) 80 100 What happens as light hits the reaction center of a photosystem? DPIP An electron from chlorophyll a is sent down the ETC, reducing NADP+ to NADPH. However, in THIS experiment, DPIP is reduced in place of NADP+. When DPIP is reduced as NADP+ normally would, it will go from blue to colorless DPIP How will the rate of photosynthesis be measured? When DPIP is reduced, less light is able to be transmitted through the chloroplasts. So by measuring the % light transmittance, the rate of photosynthesis can be deduced. 2. HYPOTHESES: How do you expect the % transmittance of light through the solutions to change in each solution of chloroplasts? What would the curves look like on each graph? Sketch them! Remember, when DPIP is reduced it will go from blue to colorless, and % transmittance will increase. 2. HYPOTHESES: How do you expect the % transmittance of light through the solutions to change in each solution of chloroplasts? What would the curves look like on each graph? Sketch them! Remember, when DPIP is reduced it will go from blue to colorless, and % transmittance will increase. Bellringer: #1. Identify a-i in the diagram. c. h. a. g. b. e. f. d. i. (2/10) 2. 3. 4. 5. 6. What replaces the electron in photosystem I? What replaces the electron in photosystem II? What is/are the product(s) of cyclic electron flow? What is/are the product(s) of noncyclic electron flow? Review EXIT. 1. 2. 3. 4. What replaces the electron in photosystem I? Electron from photosystem II What replaces the electron in photosystem II? water What is/are the product(s) of cyclic electron flow? ATP What is/are the product(s) of noncyclic electron flow? ATP & NADPH