Making energy! ATP The point is to make ATP! 2008-2009

The energy needs of life Organisms are endergonic systems What do we need energy for? synthesis building biomolecules reproduction movement active transport temperature regulation Which is to say… if you don’t eat, you die… because you run out of energy. The 2nd Law of Thermodynamics takes over!

Where do we get the energy from? Work of life is done by energy coupling use exergonic (catabolic) reactions to fuel endergonic (anabolic) reactions digestion energy + + synthesis energy + +

Build once, use many ways ATP Adenosine TriPhosphate modified nucleotide nucleotide = adenine + ribose + Pi  AMP AMP + Pi  ADP ADP + Pi  ATP adding phosphates is endergonic Marvel at the efficiency of biological systems! Build once = re-use over and over again. Start with a nucleotide and add phosphates to it to make this high energy molecule that drives the work of life. Let’s look at this molecule closer. Think about putting that Pi on the adenosine-ribose ==> EXERGONIC or ENDERGONIC? How efficient! Build once, use many ways high energy bonds

How does ATP store energy? I think he’s a bit unstable… don’t you? How does ATP store energy? P O– O –O P O– O –O P O– O –O P O– O –O P O– O –O ADP AMP ATP Each negative PO4 more difficult to add a lot of stored energy in each bond most energy stored in 3rd Pi 3rd Pi is hardest group to keep bonded to molecule Bonding of negative Pi groups is unstable spring-loaded Pi groups “pop” off easily & release energy Not a happy molecule Add 1st Pi  Kerplunk! Big negatively charged functional group Add 2nd Pi  EASY or DIFFICULT to add? DIFFICULT takes energy to add = same charges repel  Is it STABLE or UNSTABLE? UNSTABLE = 2 negatively charged functional groups not strongly bonded to each other So if it releases Pi  releases ENERGY Add 3rd Pi  MORE or LESS UNSTABLE? MORE = like an unstable currency • Hot stuff! • Doesn’t stick around • Can’t store it up • Dangerous to store = wants to give its Pi to anything Instability of its P bonds makes ATP an excellent energy donor

How does ATP transfer energy? + ATP ADP ATP  ADP releases energy ∆G = -7.3 kcal/mole Fuel other reactions Phosphorylation released Pi can transfer to other molecules destabilizing the other molecules enzyme that phosphorylates = “kinase” How does ATP transfer energy? By phosphorylating Think of the 3rd Pi as the bad boyfriend ATP tries to dump off on someone else = phosphorylating How does phosphorylating provide energy? Pi is very electronegative. Got lots of OXYGEN!! OXYGEN is very electronegative. Steals e’s from other atoms in the molecule it is bonded to. As e’s fall to electronegative atom, they release energy. Makes the other molecule “unhappy” = unstable. Starts looking for a better partner to bond to. Pi is again the bad boyfriend you want to dump. You’ve got to find someone else to give him away to. You give him away and then bond with someone new that makes you happier (monomers get together). Eventually the bad boyfriend gets dumped and goes off alone into the cytoplasm as a free agent = free Pi.

An example of Phosphorylation… Building polymers from monomers need to destabilize the monomers phosphorylate! H OH C H HO C enzyme C H OH HO O + H2O synthesis +4.2 kcal/mol + ADP C H OH “kinase” enzyme C H P + ATP Monomers  polymers Not that simple! H2O doesn’t just come off on its own You have to pull it off by phosphorylating monomers. Polymerization reactions (dehydration synthesis) involve a phosphorylation step! Where does the Pi come from? ATP It’s never that simple! -7.3 kcal/mol C H P H HO C + C H O + Pi -3.1 kcal/mol

Another example of Phosphorylation… The first steps of cellular respiration beginning the breakdown of glucose to make ATP glucose C-C-C-C-C-C Those phosphates sure make it uncomfortable around here! C H P ATP 2 hexokinase ADP 2 phosphofructokinase fructose-1,6bP P-C-C-C-C-C-C-P These are the very first steps in respiration — making ATP from glucose. Fructose-1,6-bisphosphate (F1,6bP) Dihydroxyacetone phosphate (DHAP) Glyceraldehyde-3-phosphate (G3P) 1st ATP used is like a match to light a fire… initiation energy / activation energy. The Pi makes destabilizes the glucose & gets it ready to split. DHAP P-C-C-C G3P C-C-C-P activation energy

ATP / ADP cycle Can’t store ATP ATP good energy donor, not good energy storage too reactive transfers Pi too easily only short term energy storage carbohydrates & fats are long term energy storage ATP cellular respiration 7.3 kcal/mole ADP Pi + A working muscle recycles over 10 million ATPs per second Whoa! Pass me the glucose (and O2)!

Cells spend a lot of time making ATP! The point is to make ATP! What’s the point?

Cellular Respiration Harvesting Chemical Energy ATP

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 We eat to take in the fuels to make ATP which will then be used to help us build biomolecules and grow and move and… live! heterotrophs = “fed by others” vs. autotrophs = “self-feeders”

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

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 • They are called oxidation reactions because it reflects the fact that in biological systems oxygen, which attracts electrons strongly, is the most common electron acceptor. • Oxidation & reduction reactions always occur together therefore they are referred to as “redox reactions”. • As electrons move from one atom to another they move farther away from the nucleus of the atom and therefore are at a higher potential energy state. The reduced form of a molecule has a higher level of energy than the oxidized form of a molecule. • The ability to store energy in molecules by transferring electrons to them is called reducing power, and is a basic property of living systems. loses e- gains e- oxidized reduced + – + e- e- e- oxidation reduction redox

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 p e + H – loses e- gains e- oxidized reduced oxidation reduction Energy is transferred from one molecule to another via redox reactions. C6H12O6 has been oxidized fully == each of the carbons (C) has been cleaved off and all of the hydrogens (H) have been stripped off & transferred to oxygen (O) — the most electronegative atom in living systems. This converts O2 into H2O as it is reduced. The reduced form of a molecule has a higher energy state than the oxidized form. The ability of organisms to store energy in molecules by transferring electrons to them is referred to as reducing power. The reduced form of a molecule in a biological system is the molecule which has gained a H atom, hence NAD+  NADH once reduced. soon we will meet the electron carriers NAD & FADH = when they are reduced they now have energy stored in them that can be used to do work. C6H12O6 6O2 6CO2 6H2O ATP  + oxidation H reduction e-

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 = the fuel 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 O2 is 2 oxygen atoms both looking for electrons LIGHT FIRE ==> oxidation RELEASING ENERGY But too fast for a biological system C6H12O6 6O2 6CO2 6H2O ATP  + oxidation reduction

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

Moving electrons in respiration Electron carriers move electrons by shuttling H atoms around NAD+  NADH (reduced) FAD+2  FADH2 (reduced) reducing power! P O– O –O C NH2 N+ H adenine ribose sugar phosphates NAD+ nicotinamide Vitamin B3 niacin NADH P O– O –O C NH2 N+ H H + H reduction Nicotinamide adenine dinucleotide (NAD) — and its relative nicotinamide adenine dinucleotide phosphate (NADP) which you will meet in photosynthesis — are two of the most important coenzymes in the cell. In cells, most oxidations are accomplished by the removal of hydrogen atoms. Both of these coenzymes play crucial roles in this. Nicotinamide is also known as Vitamin B3 is believed to cause improvements in energy production due to its role as a precursor of NAD (nicotinamide adenosine dinucleotide), an important molecule involved in energy metabolism. Increasing nicotinamide concentrations increase the available NAD molecules that can take part in energy metabolism, thus increasing the amount of energy available in the cell. Vitamin B3 can be found in various meats, peanuts, and sunflower seeds. Nicotinamide is the biologically active form of niacin (also known as nicotinic acid). FAD is built from riboflavin — also known as Vitamin B2. Riboflavin is a water-soluble vitamin that is found naturally in organ meats (liver, kidney, and heart) and certain plants such as almonds, mushrooms, whole grain, soybeans, and green leafy vegetables. FAD is a coenzyme critical for the metabolism of carbohydrates, fats, and proteins into energy. oxidation carries electrons as a reduced molecule

Phosphorylation SUBSTRATE LEVEL ADP receives a phosphate directly from another molecule (a phosphorylated intermediate) OXIDATIVE Oxidation of substrate releases energy; energy is used to create ATP through ATP synthase

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 C6H12O6 6O2 ATP 6H2O 6CO2  + (+ heat)

Cellular Respiration Stage 1: Glycolysis 2007-2008

What’s the point? The point is to make ATP! ATP

Where does it occur? The process of glycolysis occurs within the cell . . . but where? Glucose enters the cell through the plasma membrane (facilitated diffusion) Once it gets in the cell, glycolysis occurs within the cytoplasm Enzymes will couple with the glucose to enact all the necessary steps

In the cytosol? Why does that make evolutionary sense? Glycolysis Breaking down glucose “glyco – lysis” (splitting sugar) ancient pathway which harvests energy where energy transfer first evolved transfer energy from organic molecules to ATP still is starting point for ALL cellular respiration but it’s inefficient generate only 2 ATP for every 1 glucose occurs in cytosol In the cytosol? Why does that make evolutionary sense? glucose      pyruvate 2x 6C 3C Why does it make sense that this happens in the cytosol? Who evolved first? That’s not enough ATP for me!

Evolutionary perspective Enzymes of glycolysis are “well-conserved” Prokaryotes first cells had no organelles Anaerobic atmosphere life on Earth first evolved without free oxygen (O2) in atmosphere energy had to be captured from organic molecules in absence of O2 Prokaryotes that evolved glycolysis are ancestors of all modern life ALL cells still utilize glycolysis The enzymes of glycolysis are very similar among all organisms. The genes that code for them are highly conserved. They are a good measure for evolutionary studies. Compare eukaryotes, bacteria & archaea using glycolysis enzymes. Bacteria = 3.5 billion years ago glycolysis in cytosol = doesn’t require a membrane-bound organelle O2 = 2.7 billion years ago photosynthetic bacteria / proto-blue-green algae Eukaryotes = 1.5 billion years ago membrane-bound organelles! Processes that all life/organisms share: Protein synthesis Glycolysis DNA replication You mean we’re related? Do I have to invite them over for the holidays?

Overview 10 reactions glucose C-C-C-C-C-C fructose-1,6bP ATP 2 enzyme 10 reactions convert glucose (6C) to 2 pyruvate (3C) produces: 4 ATP & 2 NADH consumes: 2 ATP net yield: 2 ATP & 2 NADH ADP 2 enzyme fructose-1,6bP P-C-C-C-C-C-C-P enzyme enzyme enzyme DHAP P-C-C-C G3P C-C-C-P NAD+ 2 2H 2Pi enzyme 2 1st ATP used is like a match to light a fire… initiation energy / activation energy. Destabilizes glucose enough to split it in two enzyme ADP 4 2Pi enzyme ATP 4 pyruvate C-C-C DHAP = dihydroxyacetone phosphate G3P = glyceraldehyde-3-phosphate

Glycolysis summary -2 ATP 4 ATP endergonic invest some ATP exergonic ENERGY INVESTMENT -2 ATP G3P C-C-C-P exergonic harvest a little ATP & a little NADH ENERGY PAYOFF 4 ATP Glucose is a stable molecule it needs an activation energy to break it apart. phosphorylate it = Pi comes from ATP. make NADH & put it in the bank for later. like $$ in the bank net yield 2 ATP 2 NADH NET YIELD

1st half of glycolysis (5 reactions) Glucose “priming” CH2OH Glucose O 1 ATP hexokinase get glucose ready to split phosphorylate glucose molecular rearrangement split destabilized glucose ADP CH2 O P O Glucose 6-phosphate 2 phosphoglucose isomerase CH2 O P O CH2OH Fructose 6-phosphate 3 ATP phosphofructokinase P O CH2 CH2 O P ADP O Fructose 1,6-bisphosphate aldolase 4,5 H P O CH2 isomerase C O C O Dihydroxyacetone phosphate Glyceraldehyde 3 -phosphate (G3P) CHOH CH2OH CH2 O P NAD+ Pi 6 Pi NAD+ NADH glyceraldehyde 3-phosphate dehydrogenase NADH P O O CHOH 1,3-Bisphosphoglycerate (BPG) 1,3-Bisphosphoglycerate (BPG) CH2 O P

2nd half of glycolysis (5 reactions) DHAP P-C-C-C G3P C-C-C-P Energy Harvest NADH production G3P donates H oxidizes the sugar reduces NAD+ NAD+  NADH ATP production G3P    pyruvate PEP sugar donates P “substrate level phosphorylation” ADP  ATP NAD+ Pi Pi NAD+ 6 NADH NADH ADP 7 ADP O- phosphoglycerate kinase C ATP ATP CHOH 3-Phosphoglycerate (3PG) 3-Phosphoglycerate (3PG) CH2 O P 8 O- phosphoglycero- mutase C O H C O P 2-Phosphoglycerate (2PG) 2-Phosphoglycerate (2PG) CH2OH 9 O- H2O enolase H2O C O C O P Phosphoenolpyruvate (PEP) Phosphoenolpyruvate (PEP) CH2 O- ADP 10 ADP Payola! Finally some ATP! pyruvate kinase C O ATP ATP C O Pyruvate Pyruvate CH3

Substrate-level Phosphorylation In the last steps of glycolysis, where did the P come from to make ATP? the sugar substrate (PEP) H2O 9 10 Phosphoenolpyruvate (PEP) Pyruvate enolase pyruvate kinase ADP ATP CH3 O- O C P CH2 P is transferred from PEP to ADP kinase enzyme ADP  ATP ATP I get it! The Pi came directly from the substrate!

Energy accounting of glycolysis 2 ATP 2 ADP glucose      pyruvate 6C 2x 3C 4 ADP ATP 4 All that work! And that’s all I get? 2 NAD+ 2 And that’s how life subsisted for a billion years. Until a certain bacteria ”learned” how to metabolize O2; which was previously a poison. But now pyruvate is not the end of the process Pyruvate still has a lot of energy in it that has not been captured. It still has 3 carbons bonded together! There is still energy stored in those bonds. It can still be oxidized further. But glucose has so much more to give! Net gain = 2 ATP + 2 NADH some energy investment (-2 ATP) small energy return (4 ATP + 2 NADH) 1 6C sugar  2 3C sugars

Hard way to make a living! Is that all there is? Not a lot of energy… for 1 billon years+ this is how life on Earth survived no O2 = slow growth, slow reproduction only harvest 3.5% of energy stored in glucose more carbons to strip off = more energy to harvest O2 glucose     pyruvate O2 So why does glycolysis still take place? 6C 2x 3C O2 Hard way to make a living! O2 O2

raw materials  products But can’t stop there! Pi NAD+ G3P 1,3-BPG NADH DHAP 7 8 H2O 9 10 ADP ATP 3-Phosphoglycerate (3PG) 2-Phosphoglycerate (2PG) Phosphoenolpyruvate (PEP) Pyruvate NAD+ NADH Pi 6 raw materials  products Glycolysis glucose + 2ADP + 2Pi + 2 NAD+  2 pyruvate + 2ATP + 2NADH Going to run out of NAD+ without regenerating NAD+, energy production would stop! another molecule must accept H from NADH so NAD+ is freed up for another round

How is NADH recycled to NAD+? with oxygen aerobic respiration without oxygen anaerobic respiration “fermentation” Another molecule must accept H from NADH pyruvate H2O NAD+ CO2 recycle NADH NADH O2 NADH acetaldehyde acetyl-CoA NADH NAD+ NAD+ lactate lactic acid fermentation which path you use depends on who you are… Krebs cycle ethanol alcohol fermentation

Pyruvate is a branching point fermentation anaerobic respiration mitochondria Krebs cycle aerobic respiration

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

Glycolysis Overview 10 reactions glucose C-C-C-C-C-C fructose-1,6bP ATP 2 10 reactions convert glucose (6C) to 2 pyruvate (3C) produces: 4 ATP & 2 NADH consumes: 2 ATP net: 2 ATP & 2 NADH ADP 2 fructose-1,6bP P-C-C-C-C-C-C-P DHAP P-C-C-C G3P C-C-C-P NAD+ 2 2H 2Pi 1st ATP used is like a match to light a fire… initiation energy / activation energy. Destabilizes glucose enough to split it in two 2 ADP 4 2Pi ATP 4 pyruvate C-C-C

Cellular Respiration Stage 2 & 3: Oxidation of Pyruvate Krebs Cycle

Glycolysis is only the start 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 2x 6C 3C glucose      pyruvate Can’t stop at pyruvate == not enough energy produced Pyruvate still has a lot of energy in it that has not been captured. It still has 3 carbons! There is still energy stored in those bonds. pyruvate       CO2 3C 1C

Cellular respiration

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 intermembrane space inner membrane outer matrix cristae mitochondrial DNA What cells would have a lot of mitochondria?

Mitochondria – Function Oooooh! Form fits function! Mitochondria – Function Dividing mitochondria Who else divides like that? Membrane-bound proteins Enzymes & permeases bacteria! Almost all eukaryotic cells have mitochondria there may be 1 very large mitochondrion or 100s to 1000s of individual mitochondria number of mitochondria is correlated with aerobic metabolic activity more activity = more energy needed = more mitochondria What cells would have a lot of mitochondria? Active cells: • muscle cells • nerve cells What does this tell us about the evolution of eukaryotes? Endosymbiotic Theory! Advantage of highly folded inner membrane? More surface area for membrane-bound enzymes & permeases

pyruvate    acetyl CoA + CO2 Oxidation of pyruvate Pyruvate enters mitochondrial matrix 3 step oxidation process releases 2 CO2 (count the carbons!) reduces 2 NAD  2 NADH (moves e-) produces 2 acetyl CoA Acetyl CoA enters Krebs cycle [ 2x ] pyruvate    acetyl CoA + CO2 3C NAD 2C 1C Where does the CO2 go? Exhale! CO2 is fully oxidized carbon == can’t get any more energy out it CH4 is a fully reduced carbon == good fuel!!!

Pyruvate oxidized to Acetyl CoA NAD+ reduction Coenzyme A Acetyl CoA Pyruvate CO2 Release CO2 because completely oxidized…already released all energy it can release … no longer valuable to cell…. Because what’s the point? The Point is to make ATP!!! C-C C-C-C oxidation 2 x [ ] Yield = 2C sugar + NADH + CO2

Krebs cycle 1937 | 1953 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 The enzymes of glycolysis are very similar among all organisms. The genes that code for them are highly conserved. They are a good measure for evolutionary studies. Compare eukaryotes, bacteria & archaea using glycolysis enzymes. Bacteria = 3.5 billion years ago glycolysis in cytosol = doesn’t require a membrane-bound organelle O2 = 2.7 billion years ago photosynthetic bacteria / proto-blue-green algae Eukaryotes = 1.5 billion years ago membrane-bound organelles! Processes that all life/organisms share: Protein synthesis Glycolysis DNA replication

Count the carbons! x2 3C 2C 4C 6C 4C 6C 5C 4C 4C 4C pyruvate 3C 2C acetyl CoA citrate 4C 6C 4C 6C This happens twice for each glucose molecule oxidation of sugars CO2 A 2 carbon sugar went into the Krebs cycle and was taken apart completely. Two CO2 molecules were produced from that 2 carbon sugar. Glucose has now been fully oxidized! But where’s all the ATP??? x2 5C 4C CO2 4C 4C

reduction of electron carriers Count the electron carriers! CO2 pyruvate 3C 2C acetyl CoA NADH NADH citrate 4C 6C 4C 6C reduction of electron carriers This happens twice for each glucose molecule CO2 Everytime the carbons are oxidized, an NAD+ is being reduced. But wait…where’s all the ATP?? NADH x2 5C 4C FADH2 CO2 4C 4C NADH ATP

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

What’s so important about electron carriers? Electron Carriers = Hydrogen Carriers H+ 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?

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

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

What’s the point? The point is to make ATP! ATP 2006-2007

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

Cellular Respiration Stage 4: Electron Transport Chain 2006-2007

Cellular respiration

ATP accounting so far… Glycolysis  2 ATP Kreb’s cycle  2 ATP 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 lot more ATP! A working muscle recycles over 10 million ATPs per second

That sounds more like it! 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) That sounds more like it! O2

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

Electron Transport Chain Inner mitochondrial membrane Intermembrane space C Q NADH dehydrogenase cytochrome bc complex cytochrome c oxidase complex Mitochondrial matrix

Remember the Electron Carriers? glucose Krebs cycle Glycolysis G3P 2 NADH 8 NADH 2 FADH2 Time to break open the piggybank!

Electron Transport Chain Building proton gradient! NADH  NAD+ + H p e intermembrane space H+ H+ H+ inner mitochondrial membrane H  e- + H+ C Q e– e– e– H FADH2 FAD H 1 2 NADH 2H+ + O2 H2O NAD+ NADH dehydrogenase cytochrome bc complex cytochrome c oxidase complex mitochondrial matrix What powers the proton (H+) pumps?…

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 NAD+ Q C NADH H2O H+ e– 2H+ + O2 FADH2 1 2 NADH dehydrogenase cytochrome bc complex cytochrome c oxidase complex FAD H+ TA-DA!! Moving electrons do the work! Oxidation refers to the loss of electrons to any electron acceptor, not just to oxygen. Uses exergonic flow of electrons through ETC to pump H+ across membrane. H+ H+ H+ ADP + Pi ATP

electrons flow downhill to O2 But what “pulls” the electrons down the ETC? H2O Pumping H+ across membrane … what is energy to fuel that? Can’t be ATP! that would cost you what you want to make! Its like cutting off your leg to buy a new pair of shoes. :-( Flow of electrons powers pumping of H+ O2 is 2 oxygen atoms both looking for electrons O2 electrons flow downhill to O2 oxidative phosphorylation

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 ATP instead of fire! Electrons move from molecule to molecule until they combine with O & H ions to form H2O It’s like pumping water behind a dam -- if released, it can do work

“proton-motive” force We did it! H+ ADP + Pi Set up a H+ gradient Allow the protons to flow through ATP synthase Synthesizes ATP ADP + Pi  ATP ATP Are we there yet?

Chemiosmosis links the Electron Transport Chain to ATP synthesis 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 Chemiosmosis is the diffusion of ions across a membrane. More specifically, it relates to the generation of ATP by the movement of hydrogen ions across a membrane. Hydrogen ions (protons) will diffuse from an area of high proton concentration to an area of lower proton concentration. Peter Mitchell proposed that an electrochemical concentration gradient of protons across a membrane could be harnessed to make ATP. He likened this process to osmosis, the diffusion of water across a membrane, which is why it is called chemiosmosis. So that’s the point!

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

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

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

Summary of cellular respiration C6H12O6 6O2 6CO2 6H2O ~40 ATP  + 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? Where did the glucose come from? from food eaten Where did the O2 come from? breathed in Where did the CO2 come from? oxidized carbons cleaved off of the sugars (Krebs Cycle) Where did the CO2 go? exhaled Where did the H2O come from? from O2 after it accepts electrons in ETC Where did the ATP come from? mostly from ETC What else is produced that is not listed in this equation? NAD, FAD, heat!

cytochrome c oxidase complex NAD+ Q C NADH H2O H+ e– 2H+ + O2 FADH2 1 2 NADH dehydrogenase cytochrome bc complex cytochrome c oxidase complex FAD 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! What if you have a chemical that punches holes in the inner mitochondrial membrane?

Fermentation (anaerobic) Bacteria, yeast 1C 3C 2C pyruvate  ethanol + CO2 NADH NAD+ back to glycolysis beer, wine, bread Animals, some fungi Count the carbons!! Lactic acid is not a dead end like ethanol. Once you have O2 again, lactate is converted back to pyruvate by the liver and fed to the Kreb’s cycle. pyruvate  lactic acid 3C NADH NAD+ back to glycolysis cheese, anaerobic exercise (no O2)

Alcohol Fermentation pyruvate  ethanol + CO2 Dead end process bacteria yeast Alcohol Fermentation recycle NADH 1C 3C 2C pyruvate  ethanol + CO2 NADH NAD+ back to glycolysis Dead end process at ~12% ethanol, kills yeast can’t reverse the reaction Count the carbons!

Lactic Acid Fermentation animals some fungi recycle NADH Lactic Acid Fermentation O2 pyruvate  lactic acid 3C  NADH NAD+ back to glycolysis Reversible process once O2 is available, lactate is converted back to pyruvate by the liver Count the carbons!

What’s the point? The point is to make ATP! ATP 2006-2007

Heat O2 ATP H2O NAD+ NADH CO2 Glucose ATP Pyruvate MITOCHONDRION Electron Transport System O2 ATP H2O NAD+ NADH CO2 citric acid cycle Glucose ATP Pyruvate MITOCHONDRION