Section 4. Fuel oxidation, generation of ATP Section 4. Overview of Fuel oxidation, ATP generation: Physiological processes require energy transfer from chemical bonds in food: Electrochemical gradient Movement of muscle Biosynthesis of complex molecules 3 phases: Oxidation of fuels (carbs, fats, protein) Conversion of energy to ~PO4 of ATP Utilization of ATP to drive energy-requiring reactions Fig.iv.1

Fuel oxidation overview - respiration Phase 1: energy (e-) from fuel transfer to NAD+ and FAD; Acetyl CoA, TCA intermediates are central compounds Phase 2: electron transport chain convert e- to ATP; membrane proton gradient drives ATP synthase Phase 3: ATP powers processes Fig. iv.2

Respiration occurs in mitochondria Most enzymes in matrix Inner surface has e- transport chain ATP synthase ATP transported through inner membrane, diffuses through outer Some enzymes encoded by mitochondrion genome, most by nuclear genes Fig. iv.3

Glucose is universal fuel for every cell Glycolysis is universal fuel: 1 glucose -> 2 pyruvate + 2 NADH + 2 ATP Aerobic path: Continued oxidation Acetyl CoA -> TCA, NADH, FAD(2H) -> e- transport chain Lots of ATP Anaerobic: fermentation: ‘anaerobic glycolysis’ Oxidation of NADH to NAD+ Wasteful reduction of pyruvate to lactate in muscles to ethanol, CO2 by yeast Fig. iv.4

Chapt. 19 Cellular bioenergetics of ATP, O2 Ch. 19 Cellular bioenergetics Student Learning Outcomes: Explain the ATP-ADP cycle Describe how chemical bond energy of fuels can do cellular work through ~PO4 bond of ATP Explain how NADH, FAD(2H) coenzymes carry electrons to electron transport chain Describe how ATP synthesis is endergonic (requires energy) Describe how ATP hydrolysis (exergonic) powers biosynthesis, movement, transport

Fuel oxidation makes ATP Cellular Bioenergetics of ATP and O2: Chemical bond energy of fuels transforms to physiological responses necessary for life Fuel oxidation generates ATP ATP hydrolysis provides energy for most work High energy bonds of ATP: Energy currency of cell Fig. 19.1

High energy phosphate bond of ATP: Strained phosphoanhydride bond DG0’ -7.3 kcal/mol standard conditions Hydrolysis of ATP to ADP + Pi transfers PO4 to metabolic intermediate or protein, for next step Fig. 19.2

Thermodynamics states what is possible: Thermodynamics brief Thermodynamics states what is possible: DG = change in Gibbs free energy of reaction: DG = DG0 + RT ln [P]/[S] (R = gas const; T = temp oK) DG0 = DG at standard conditions of1 M substrate & product and proceeding to equilibrium) DG0’ = DG0 under standard conditions of [H2O] = 55.5 M, pH 7.0, and 25oC [37oC not much different] Concentrations of substrate(s) and products(s): At equilibrium, DG = 0, therefore DG0’ = -RT lnKeq’ = -RT ln[P]/[S]

Thermodynamics states what is possible: Thermodynamics brief Thermodynamics states what is possible: Exergonic reactions give off energy (DG0’ < 0) typically catabolic Endergonic reactions require energy (DG0’ > 0) typically anabolic Unfavorable reactions are coupled to favorable reactions Hydrolysis of ATP is very favorable Additive DG0’ values determine overall direction

C. Exogonic, endogonic reactions Phosphoglucomutase converts G6P to/from G1P: G6P to glycolysis G1P to glycogen synthesis Equilibrium favors G6P Exergonic reactions give off energy (DG0’ < 0) Endergonic reactions require energy (DG0’ > 0) Fig. 19.3

III. Energy transformation for mechanical work ATP hydrolysis can power muscle movement: Myosin ATPase hydrolyzes ATP, changes shape ADP form changes shape back, moves along Actin was activated by Ca2+ Fig. 19.4

Active transport: ATP hydrolysis moves molecules: ATP powers transport Active transport: ATP hydrolysis moves molecules: Na+, K+ ATPase sets up ion gradient; bring in items Vesicle ATPases pump protons into lysosome Ca2+-ATPases pump Ca2+ into ER, out of cell Fig. 10.6

III. ATP powers biochemical work ATP powers biochemical work, synthesis: Anabolic paths require energy: DGo’ additive Couple synthesis to ATP hydrolysis: Phosphoryl transfer reactions Activated intermediate Ex. Table 19.3: glucose + Pi -> glucose 6-P + H2O + 3.3 kcal/mol ATP + H2O -> ADP + Pi - 7.3 kcal/mol Sum: glucose + ATP -> glucose 6-P + ADP -4.0 Also Glucose -> G-1-P will be -2.35 kcal/mol overall: hydrolysis of ATP, through G-6-P to G-1-P

Activated intermediates in glycogen synthesis Glycogen synthesis needs 3 ~P: Phosphoryl transfer to G6P Activated intermediate with UDP covalently linked Fig. 19.5 Fig. 19.6

DG depends on substrate, product concentrations DG = DG0 + RT ln [P]/[S] Cells do not have 1M concentrations High substrate can drive reactions with positive DG0’ Low product (removal) can drive reactions with positive DG0’ Ex., even though equilibrium (DG0’= +1.6 kcal/mol) favors G6P: G1P in a ratio 94/6, If G1P is being removed (as glycogen synthesis), then equilibrium shifts ex. If ratio 94/3, then DG = -0.41 favorable

Activated intermediates with ~bonds Other compounds have high-energy bonds to aid biochemical work: (equivalent to ATP) UTP, CTP and GTP also (made from ATP + NDP): UTP for sugar biosyn, GTP for protein, CTP for lipids Some other compounds: Creatine PO4 energy reserve muscle, nerve, sperm Glycolysis Ac CoA TCA cycle Fig. 19.7

V. Energy from fuel oxidation Energy transfer from fuels through oxidative phosphorylation in mitochondrion: NADH, FAD(2H) transfer e- to O2 Stepwise process through protein carriers Proton gradient created e- to O2 -> H2O ATP synthase makes ATP lets in H+ Fig. 19.8

Oxidation: reduction reactions: Electron donor gets oxidized; recipient is reduced LEO GER: Loss Electrons = oxidation; gain electrons is reduction use coenzyme e- carriers Fig. 19.9 NADH Fig. 19.10 FAD(2H)

Redox potentials indicate energetic possibility: Energy tower; combine half reactions for overall: Ex. Table 19.4: ½ O2 + 2H+ + 2e- -> H2O E0’ 0.816 NAD+ + 2H+ + 2e- -> NADH + H+ -0.320 Combine both reactions (turn NADH -> NAD+) = 0.320 Total 1.136 (very big) = -53 kcal/mol FAD(2H) gives less, since its only +0.20 (FAD(2H) -> FAD

Calorie content of fuels reflects oxidation state C-H and C-C bonds will be oxidized: Glucose has many C-OH already: 4 kcal/g Fatty acids very reduced: 9 kcal/g Cholesterol no calories: not oxidized in reactions giving NADH

Anaerobic glycolysis” = fermentation In absence of O2, cell does wasteful recycling: NADH oxidized to NAD+ (lose potential ATP) pyruvate reduced to lactate glycolysis can continue with new NAD+ yeast makes ethanol, CO2 from pyruvate bacteria make diverse acids, other products Fig. 19.11

Oxidation not for ATP generation Most O2 used in electron transport chain. Some enzymes use O2 for substrate oxidation, not for ATP generation: Oxidases transfer e- to O2 [Cytochrome oxidase in electron transport chain] Peroxidases in peroxisome Oxygenases transfer e- and O2 to substrate Form H2O and S-OH Hydroxylases (eg. Phe -> Tyr) Fig. 19.12

Energy expenditure reflects oxygen consumption: Most O2 is used VII Energy balance Energy expenditure reflects oxygen consumption: Most O2 is used by ATPases Fig. 19.14

Portion of food metabolized is related to energy use: Energy balance Portion of food metabolized is related to energy use: Basal metabolic rate Thermogenesis Physical activity Storage of excess “If you eat to much and don’t exercise, you will get fat” (summarizes ATP-ADP cycle)