biochemistry for nursing Krebs and ETC.pptx

overview of TCA and ETC

1. TCA CYCLE DR. CHIKONDE

2. Definition The Krebs cycle, also known as the citric acid cycle or TCA cycle is a series of reactions that take place in the mitochondria resulting in oxidation of acetyl CoA to release carbon dioxide and hydrogen atoms that later lead to the formation of water. This cycle is termed the citric acid cycle as the first metabolic intermediate formed in the cycle is citric acid. This cycle is also termed tricarboxylic acid (TCA).

3. This cycle only occurs under aerobic conditions as energy-rich molecules like NAD +  and FAD can only be retrieved from their reduced form once they transfer electrons to molecular oxygen. The citric acid cycle is the final common pathway for the oxidation of all biomolecules; proteins, fatty acids, carbohydrates. Molecules from other cycles and pathways enter this cycle through Acetyl CoA. The citric acid cycle is a cyclic sequence of reactions formed of 8 enzyme-mediated reactions. This cycle is also particularly important as it provides electrons/ high-energy molecules to the electron transport chain for the production of ATPs and water.

4. Pyruvate formed at the end of  glycolysis  is first oxidized into Acetyl CoA which then enters the citric acid cycle.

5. Krebs cycle Location The citric acid cycle in eukaryotes takes place in the mitochondria while in prokaryotes, it takes place in the cytoplasm. The pyruvate formed in the cytoplasm (from glycolysis) is brought into the mitochondria where further reactions take place. The different enzymes involved in the citric acid cycle are located either in the inner membrane or in the matrix space of the mitochondria.

6. Krebs cycle Equation/ Reaction The overall reaction/ equation of the citric acid cycle is: Acetyl CoA + 3 NAD +  + 1 FAD + 1 ADP + 1 P i     →   2 CO 2  + 3 NADH + 3 H +  + 1 FADH 2  + 1 ATP In words, the equation is written as: Acetyl CoA + Nicotinamide adenine dinucleotide + Flavin adenine dinucleotide + Adenosine diphosphate + Phosphate   →   Pyruvate + Water + Adenosine triphosphate + Nicotinamide adenine dinucleotide + Hydrogen ions

7. ENZYMES Citrate synthase Aconitase Isocitrate dehydrogenase α-ketoglutarate Succinyl-CoA synthetase Succinate dehydrogenase Fumarase Malate dehydrogenase In eukaryotic cells, the enzymes that catalyze the reactions of the citric acid cycle are present in the matrix of the mitochondria except for succinate dehydrogenase and aconitase, which are present in the inner mitochondrial membrane. One common characteristic in all the enzymes involved in the citric acid cycle is that nearly all of them require Mg 2+

8. Krebs cycle Steps The oxidative decarboxylation of pyruvate forms a link between glycolysis and the citric acid cycle. In this process, the pyruvate derived from glycolysis is oxidatively decarboxylated to acetyl CoA and CO 2  catalyzed by the pyruvate dehydrogenase complex in the mitochondrial matrix in eukaryotes and in the cytoplasm of the prokaryotes. From one molecule of glucose, two molecules of pyruvate are formed, each of which forms one acetyl CoA along with one NADH by the end of the pyruvate oxidation. The acetyl CoA formed from pyruvate oxidation, fatty acid metabolism, and amino acid pathway then enter the citric acid cycle.

9. Step 1: Condensation of acetyl CoA with oxaloacetate The first step of the citric acid cycle is the joining of the four-carbon compound oxaloacetate (OAA) and a two-carbon compound acetyl CoA. The oxaloacetate reacts with the acetyl group of the acetyl CoA and water, resulting in the formation of a six-carbon compound citric acid, CoA. The reaction is catalyzed by the enzyme citrate synthase that condenses the methyl group of acetyl CoA and the carbonyl group of oxaloacetate resulting in citryl-CoA which is later cleaved to free coenzyme A and to form citrate.

10. Step 2: Isomerization of citrate into isocitrate Now, for further metabolism, citrate is converted into isocitrate through the formation of intermediate cis-aconitase. This reaction is a reversible reaction catalyzed by the enzyme (aconitase). This reaction takes place by a two-step process where the first step involves dehydration of citrate to cis-aconitase, followed by the second step involving rehydration of cis-aconitase into isocitrate.

11. Step 3: Oxidative decarboxylations of isocitrate The third step of the citric acid cycle is the first of the four oxidation-reduction reactions in this cycle. Isocitrate is oxidatively decarboxylated to form a five-carbon compound, α-ketoglutarate catalyzed by the enzyme isocitrate dehydrogenase. This reaction, like the second reaction, is a two-step reaction. In the first step, isocitrate is dehydrogenated to oxalosuccinate while the second step involves the decarboxylation of oxalosuccinate to α-ketoglutarate. Both the reactions are irreversible and catalyzed by the same enzyme. The first step, however, results in the formation of NADH while the second step involves the release of CO 2 .

12. Step 4: Oxidative decarboxylation of α-ketoglutarate This step is another one of the oxidation-reduction reactions where α-ketoglutarate is oxidatively decarboxylated to form a four-carbon compound, succinyl-CoA, and CO 2 . The reaction irreversible and catalyzed by the enzyme complex α-ketoglutarate dehydrogenase found in the mitochondrial space. This reaction is similar to the oxidative decarboxylation of pyruvate involving the reduction of NAD +  into NADH.

13. Step 5: Conversion of succinyl-CoA into succinate In the next step, succinyl-CoA undergoes an energy-conserving reaction in which succinyl-CoA is cleaved to form succinate. This reaction is accompanied by phosphorylation of guanosine diphosphate (GDP) to guanosine triphosphate (GTP). The GTP thus formed then readily transfers its terminal phosphate group to ADP forming an ATP molecule. The reaction is catalyzed by the enzyme, succinyl-CoA synthase.

14. Step 6: Dehydration of succinate to fumarate Here, the succinate formed from succinyl-CoA is dehydrogenated to fumarate catalyzed by the enzyme complex succinate dehydrogenase found in the intramitochondrial space. This is the only dehydrogenation step in the citric acid cycle in which NAD +  doesn’t participate. Instead, another high-energy electron carrier, flavin adenine dinucleotide (FAD) acts as the hydrogen acceptor resulting in the formation of FADH 2 . The FADH 2  then enters the electron transport chain via the complex II transferring the electrons to ubiquinone, finally forming 2ATPs.

15. Step 7: Hydration of fumarate to malate The fumarate is reversibly hydrated to form L-malate in the presence of the enzyme fumarate hydratase. As it is a reversible reaction, the formation of L-malate involves hydration, whereas the formation of fumarate involves dehydration.

16. Step 8: Dehydrogenation of L-malate to oxaloacetate The last step of the citric acid cycle is also an oxidation-reduction reaction where L-malate is dehydrogenated to oxaloacetate in the presence of L-malate dehydrogenase, which is present in the mitochondrial matrix. This is a reversible reaction involving oxidation of L-malate and reduction of NAD +  into NADH. Oxaloacetate thus formed, allows the repetition of the cycle and NADH formed participates in the oxidative phosphorylation. This reaction completes the cycle.

17. Products of KREBS cycle

18. At each turn of the cycle, 3 NADH, 1 FADH2, 1 GTP (or ATP), 2 CO2 Note:  One NADH are formed from a molecule of pyruvate in the oxidative decarboxylation of pyruvate to Acetyl CoA.

19. THANK YOU

20. ELECTRON TRANSPORT CHAIN DR CHIKONDE

21. 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 ! What’s the point? A working muscle recycles over 10 million ATPs per second There’s got to be a better way!

22. There is a better way! Electron Transport Chain series of molecules 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 ~34 ATP from 1 glucose ! only in presence of O 2 ( aerobic respiration ) O 2 That sounds more like it !

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

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

25. G3P Glycolysis Krebs cycle 8 NADH 2 FADH 2 Remember the Electron Carriers? 4 NADH Time to break open the bank ! glucose

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

27. Electrons flow downhill Electrons move in steps from carrier to carrier downhill to O 2 each carrier more electronegative controlled oxidation controlled release of energy make ATP instead of fire !

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

29. ELECTRON TRANSPORT CHAIN(ETC) ETC couple a chemical reaction b/w an electron donor and electron acceptor to the transfer of H+ ions across a membrane, through a set of mediating biochemical reactions These H+ ions are used to produce ATP ETC used for extracting energy from sunlight(photosynthesis) and from redox reactions such as the oxidation of sugar (respiration)

30. CELLULLAR RESPIRATION

31. COMPONENTS OF ETC NAD & F lavoprotein :H-carriers in celluiar respiration Non heme metalloprotein (Fe-S- Protein ): iron cycles between 3+ and 2+ states. Ubiquinone or CoQ: region serves as an anchor to inner mitochondrial membrane. Cytochromes : Electron-transfer proteins that contain a heme prosthetic group

32. Composition of the Electron Transport Chain Four large protein complexes. Complex I - NADH-Coenzyme Q reductase Complex II - Succinate-Coenzyme Q reductase Complex III - Cytochrome c reductase Complex IV - Cytochrome c oxidase Many of the components are proteins with prosthetic groups to move electrons.

33. Complex I(NADH Dehydrogenase) Electrons pass from NADH 🡪 FMN 🡪 Fe-S cluster 🡪 ubiquinone (flavin mononucleotide) ( coenzyme Q)

34. Complex II(succinate dehydrogenase) Entry point for FADH 2 . Succinate dehydrogenase ( from the citric acid cycle ) directs transfer of electrons from succinate to CoQ via FADH 2 . Acyl-CoA dehydrogenase (from β -oxidation of fatty acids) also transfers electrons to CoQ via FADH 2 .

35. Complex III (cytochromes b, c1 and c ) Electron transfer from ubiquinol to cytochrome c. cytochrome c heme prosthetic group

36. Complex IV Combination of cytochromes a and a 3 , 10 protein subunits, 2 types of prosthetic groups: 2 heme and 2 Cu. Electrons are delivered from cytochromes a and a3 to O2. Several chemicals can inhibit the pathway at different locations. Cyanide and CO can block e transport between a/a3 and O2.

37. Flow of electrons c y t c Q C o m p l e x I C o m p l e x I I C o m p l e x I I I C o m p l e x I V - . 4 - . 2 . . 2 . 4 . 6 . 8 1 . N A D H N A D + s u c c i n a t e f u m a r a t e H O 2 1 / 2 O + 2 H 2 + P a t h o f E l e c t r o n s (FADH2) Energy is not released at once, but in incremental amounts at each step.

38. Inner mitochondrial membrane Outer mitochondrial membrane H + H + H + H + H + H + H + H + H + H + H + H + H + H + H + ADP + P i ATP Electron Transport Chain ATP synthase complex

39. FUNCTIONING OF ETC In ETC , the constituent molecule are arranged in the order of 🡺increasing redox potential 🡺Decreasing e- pressure 🡺Diminishing free energy level so along the chain there would be a step by step flow of e- from most –ve initial donor to most +ve terminal acceptor(O2) e- entering to ETC are energy rich As the flow down , they loss free energy Much of this energy get conserved in ATP

40. 🡺in ETC enzyme bound H is used as the fuel for energy generation 🡺available from the dehydrogenation reactions of substrate oxidation 🡺from H-donating substrates ,H-atoms are collected by dehydrogenase , and supplied to ETC , through NAD &FAD 🡺They are serve as the acceptor , carrier & donor of H. 🡺The H atoms soon donate their e- to ETC 🡺These H+ ions soon escape to the aquas medium

41. NAD linked dehydrogenase remove 2 H-atoms from the substrate 1-transferred to NAD as hydride ion 1-appers in the medium as H+ 🡺reduced substrate + NAD + oxidized substrate + NADH+H 🡺FAD-linked dehydrogenase remove 2 H atoms from the substrate 🡺in most case ,both of them will be accepted by FAD ,yielding FADH2 🡺reduced substrate + FAD oxidized subsrtate+FADH2

42. 🡺from the co-enzyme channel , electrons are funneled to molecular oxygen though chain of e- carrier 🡺they include flavoproteins , Fe-S,UQ, Co Q & cytochromes 🡺in respiratory chain e- are transferred in 3 ways 1.As e- s , 2.as H-atoms,3. as hydride ions which bears two electrons 🡺during the flow of e-,every members of ETC get Alternately reduced and oxidized in cyclic manner 🡺in this process ,iron atoms oscillates between Fe2+ &Fe3+ states

43. 🡺at the initial end , this redox reaction chain is linked to dehydrogenation reaction , and at its terminal end to molecular oxygen 🡺at the same time , some of its intermediate reactions are coupled with phosphorylation of ADP 🡺dehydrogenation reactions maintain a steady flow of H & Phosphorylation reaction ensure a constant synthesis of ATP 🡺 The coupling of redox reactions of the respiratory chain with the phosphorylation of ADP constitutes a reaction complex known as oxidative phosphorylation

44. Oxidative phosphorylation The electron-transport chain moves electrons from NADH and FADH 2 to O 2 . In the mean time, ADP is phosphorylated to ATP. The two processes are dependent on each other. ATP cannot be synthesized unless there is energy from electron transport (ΔG o ’= +31 kj/mol). Electrons do not flow to O2, unless there is need for ATP.

45.   3 ATP are generated when two electrons are transported from NADH to O2. The oxidation of FADH2 only produces 2 ATP.