1. Mitochondrion and Chloroplast Ultrastructure

3. Discovery of Mitochondria
Kollicker (1850) teased mitochondria from striated muscle
Altmann (1890) used special stains to identify these granules and named them bioblasts
Warburg (1910) used low-speed centrifugation to isolate mitochondria and showed it contained enzymes for catalysing oxidative reactions
1950 Lehninger, Green and others clearly showed that fatty acid oxidation and oxidative phosphorylation were properties of mitochondria

4. Structure of Mitochondria
The number and distribution of mitochondria in a cell are closely related to the activity of the cell. Eg cells undergoing movement may display increased numbers of mitochondria
Most are ovoid bodies with a diameter of between 0.5 – 1.0 micrometers and a length of up to 7 micrometers

5. Mitochondria Ultrastructure

6. Structure of Mitochondria
Outer membrane- contains special pores which make it freely permeable to most small molecules and ions
Matrix-gel-like solution which contains enzymes, coenzymes responsible for fatty acid oxidation etc.
Inner membrane-is specialized highly convoluted membrane which is impermebale to most small ions like H+, ATP, ADP
Specialized transport systems are necessary to move ions across the membrane

7. ATP SYNTHETASE COMPLEXES
These are protein complexes referred to as inner membrane particles and are attached to the inner surface of the inner mitochondrial membrane
They appear as spheres that protrude into the mitochondrial matrix

8. Matrix of the Mitochondria
Gel-like solution contains enzymes responsible for the oxidation of pyruvate, amino acids, fatty acids and those of the TCA cycle
The synthesis of urea and heme occur partially in the matrix
Contains coenzymes NAD+ and FAD (the oxidized forms of the 2 coenzymes that are required as hydrogen acceptors
Contains ADP and Pi which combine to produce ATP

9. Electron Transport Chain
Located in the inner mitochondrial membrane
Energy-rich molecules are metabolized by a series of oxidation reactions to yield CO2 andH2O
The metabolic intermediates donate electrons to specialized coenzymes NAD+ and FAD to form energy rich NADH and FADH2. These in turn donate a pair of their electrons to a special set of electron carriers collectively called the ELECTRON TRANSPORT CHAIN
As electrons move down the chain they lose free energy, part of this energy is used in the production of ATP. This process is called OXIDATIVE PHOSPHORYLATION

10. Organization of the Chain
The inner mitochondrial membrane can be disrupted into five separate enzyme complexes
Complex IComplex IV each contain the electron transport chain
Complex V catalyzes ATP synthesis
Each complex Accepts OR Donates electrons to the next carrier in the chain
The end result is a combination of protons with oxygen to form water
This requirement for oxygen makes the electron transport chain O2 utilization the greatest in the body

12. Reactions of the Electron Transport Chain
All members of the chain excepting Coenzyme Q (ubiquinone) are protein
May be enzymes eg dehydrogenases
May contain iron as a part of an iron-sulphur center
May be coordinated with an porphyrin ring like the cytochromes
May contain copper eg cytochromes a+a3

13. Reactions of the Electron Transport Chain
Formation of NADH
NADH dehydrogenase enzyme complex
Coenzyme Q
Cytochromes B and C
Cytochrome a+a3

14. 1. Formation of NADH
NAD+ is reduced to NADH by dehydrogenases which remove 2 Hydrogen atoms
Both electrons but one proton (H- hydride ion)are transferred to the NAD+ to form NADH

15. 2. NADH Dehydrogenase
The free proton plus the hydride ion carried on NADH are transferred to NADH dehydrogenase (NADH-Q reductase)
Enzyme complex embedded on the inner membrane
FMN, a coenzyme accepts the 2 hydrogen atoms and forms FMNH2

16. Coenzyme Q
Coenzyme Q can accept both hydrogen atoms from FMNH2 and FADH2. FADH2 is produced by succinate dehydrogenase and acyl CoA dehydrogenase

17. Cytochromes b,c and a+a3
Cytochromes contain a heme group and a porphyrin ring
Electrons are passed down the chain from coenzyme Q to cyochromes b,c and a+a3
Cytochrome a+a3 is the only electron carrier in which the heme iron has a free ligand and can react directly with molecular oxygen
Transported electrons bind with oxygen to produce water

18. ELECTRON TRANSPORT CHAIN

20. Inhibitors of electron transfer
Rotenone and amytal block electron transfer in the NADH-Q reductase
Antimycin A blocks from cytochrome b reductase
CN-, N3- and CO block cytochrome oxidase by binding with the heme at a3

21. OXIDATIVE PHOSPHORYLATION

22. ATP Formation
Produced in the cytosol during glycolysis in which sugars are catabolized
May be produced within chloroplasts by the utilisation of light energy
May be produced within the mitochondria present in all plant and animal cells by the oxidation of elementary substrates

23. Oxidative Phosphorylation
The process by which ATP is formed as a result of the transfer of electrons from NADH or FADH2 to oxygen by a series of electron carriers
THIS IS THE MAJOR SOURCE OF ATP
Generates 26 of the 30 molecules of ATP formed when glucose is completely oxidized to CO2 and H2O

24. Electron motive force is converted to proton-motive force and then into phosphoryl potential
Oxidation and phosphorylation are coupled

25. NADH +1/2 O2 +H+ -H2O +NAD+ ADP+Pi+H+-ATP+ H2O ΔG°’ = -52.6 kcal/mol

26. The Proton Pump
Electron transport is coupled with the transport of protons across the inner mitochondrial membrane from the matrix to the inner membrane space
This creates an electron gradient with more +ve charges outside the membrane
A pH gradient is also created, the outside of the membrane is at a lower potential than the inside
The energy generated is sufficient to drive ATP synthesis

27. Chemiosmotic Hypothesis
After the protons have been transferred to the cytosolic side of the inner mitochondrial membrane they reenter the mitochondrial matrix by passing through a channel in the ATP synthetase molecule
Thus forming ATP from ADP and Pi
ATPase spheres can be seen on the inner mitochondrial membrane and are referred to as F1
F1 unit catalyzes the synthesis of ATP

29. The other major unit of ATP synthase is Fo,
A hydrophobic segment which spans the inner membrane
Is the protein channel of the complex
The stalk between F1 and F0 is sensitive to oligomycin

30. ATP Synthesis
The enzyme complex ATP synthetase (ATPase) synthesizes ATP utilising the energy generated by the proton motive force of the electron transport chain
Enzyme bound ATP forms without the presence of the proton motive force
The proton gradient releases ATP from the synthase complex
Catalyzes the formation of ATP from ADP and orthophosphate (Pi)

31. Electrons from cytosolic NADH enter mitochondria by shuttle
Electrons from NADH rather than NADH itself are carried across the mitochondrial membrane
Glycerol phosphate shuttle
Malate aspartate shuttle

32. The entry of ADP into Mitochondria is coupled to the exit of ATP by the ATP-ADP translocase
ADP only enters the mitochondrial matrix if ATP exits and vice versa
Mitochondrial transporters for metabolites have a common tripartite motif

34. THE Rate of oxidative phosphorylation is dependent on the need for ATP
This is determined by the level of ADP
The rate of O2 consumption increases when ADP is added then returns to its original value when ATP has been synthesized
Electrons do not flow from fuel molecules unless ATP needs to be synthesized

35. ATP yield Glycolysis 2 Citric acid cycle 2
Oxidative phosphorylation 26
TOTAL

36. CHLOROPLAST STRUCTURE

37. ALL free energy for biological systems arises from solar energy by a process known as PHOTOSYNTHESIS
Photosynthesis takes place in chloroplasts
Chloroplast are 5 micrometers long
Like mitochondria they contain an outer and an inner membrane and an intermembrane space

38. The inner membrane surrounds a stroma, containing soluble enzymes
Membranous structures called thylakoids (flattened discs)
A pile of thylakoids is called a granum
Thylakoid membranes contain energy transducing machinery: the light harvesting proteins, reaction centers electron transport chains and ATP synthase

39. The outer membrane is permeable to small ions and molecules
The stroma contains soluble enzymes that utilise NADPH and ATP synthesized by the thylakoids to produce sugar

40. Chlorophyll The principal photoreceptor
Is a magnesium porphyrin whereas heme is an iron porphyrin
Have very strong absorption bands because they contain networks of alternating single and double bonds called polyenes

41. REACTION CENTER
Photosynthesis can be separated into light and dark reactions
The light reactions generate NADPH and ATP
The dark reaction use NADPH and ATP to reduce CO2

42. Transmembrane proton gradient
Electon flow from PII to PI as well as within the system generates a transmembrane proton gradient that drives ATP formation
Photosystem II transfers electrons from water to plastoquinone
Plastiquinone resembles ubiquinone (CoQ)

43. D1 and D2
Phtosystem II is formed by D1 and D2 a pair of subunits that span the thylakoid membrane
D1 and D2 contain the reaction center and the electron transfer chain

44. Electron Transfer PI Electronic Excitation of P680 Binds to pheophytin
Binds to plastoquinone at the Qa site
Reduces plastoquinone to quinone
Binds to Qb site
Electron flows from H2O to QH2
Formation of a proton gradient as the electron flows through cytochrome bf complex catalyses the transfer of electrons plastoquinol to (QH2) to plastocyanin and concomitantly pumps protons across the thylakoid membrane

45. Electron transfer PI
An electron is transferred from P700 (reaction site of PI)
P700 gets excited
The electron is transferred from P700 to Ao ( a monomeric acceptor chlorophyll)
AoAo- and P700+ (most potent reductant in biological systems
P700+ captures an electron from plastocyanin P700
Ao- is transferred to A1 a quinone and hen to Fx an iron-sulphur cluster
FxFerredoxin
Net rxn PI is PC (Cu+)+ ferredoxin (oxidized)-->PC (Cu++) +ferredoxin (reduced)

46. NADPH Formation
High potential electrons of ferredoxin are transferred to NADP+ to form NADPH
This rxn is catalyzed by a flavoprotein ferredoxin-NADP+ reductase
The net rxn of PII is
2H2O + 2 NADP+  O2 +2NADPH + 2H+
Light causes electrons to flow from H2O to NADPH and leads to the generation of a proton-motive force

47. ATP synthesis
Driven by a proton-motive force in both photphosphorylation and oxidative phosphorylation
ATP synthase is very similar to mitochondria the complex is called CFo-CF1
Recurring motifs and mechanisms in photosynthetic reaction centers

48. The Reaction center
P680 is a reaction center

52. CITRIC ACID CYCLE