In The name of Almighty Allah, The Merciful, The Beneficent

Assalam u Alykum WRWB

Electron transport and Oxidative Phosphorylation

Objectives
1. 2. Define reducing equivalents and list their sources and fates. List the components of electron transport chain and describe the process of electron transport Define oxidative phosphorylation and list their products

3.

4.
5.

Explain the importance of oxidative phosphorylation

Define energy, energy currency (ATP) and energy storage (creatine phosphate) ad describe the functions of Creatine phosphate and ATP.

 Biological oxidations are catalyzed by intracellular enzymes. The purpose of oxidation is to obtain energy. Electron Transport: Electrons carried by reduced coenzymes (NADH or FADH2) are passed sequentially through a chain of proteins and coenzymes (so called electron transport chain)to O2 . Oxidative Phosphorylation: Coupling e- Transport (Oxidation) and ATP synthesis (Phosphorylation) . It all happens in mitochondrion or at the inner mitochondrial membrane (eukaryotic cells)

Energy and Free energy

All biochemical processes (reactions) involve the energy transactions
Exergonic reactions release energy Endergonic reactions- consume energy The actual energy available to do work (utilizable) is known as free energy.

Reducing equivalents
The free energy stored in food stuffs is extracted in the form of reducing equivalents. The reducing equivalents are defined as a proton plus an electron, H+ + e The reducing equivalents are transferred to ETC via different carriers like NAD, FAD, FMN etc ATP is synthesized by oxidative Phosphorylation.

Reducing equivalents
Are transported Via Conjugate oxidant/reductant pairs NAD-NADH FAD-FADH2 FMN-FMNH2 Redox Potential (E0): is the quantitative measure of tendency of a redox pair to loose or gain electrons.

A more negative E0 means a greater tendency to loose electrons, and a positive E0 means a greater tendency to gain electrons.

Electron Transport (ETC)

the mitochondrion contained the enzymes responsible for electron transport and oxidative phosphorylation

In inner membrane knobs

Impermeable to ions and most other compounds

Electron Carriers
The transfer of electrons is not directly to oxygen but through coenzymes There are 2 sites of entry NAD+ for electrons into the FMN electron transport chain: NAD+ or FAD
FeS FAD FeS ubiquinone Cyt b ubiquinone FeS Cyt c1 Cyt c Cyt a Cyt a3 1/2 O2

Both are coenzymes for dehydrogenase enzymes

 Coenzyme Q (CoQ, Q or ubiquinone) is lipidsoluble. It dissolves in the hydrocarbon core of a membrane. the only electron carrier not bound to a protein.
it can accept/donate 1 or 2 e-. Q can mediate etransfer between 2 e- that transfer and 1 e- carriers

Cytochromes
NAD+ FMN FeS FAD FeS ubiquinone Cyt b ubiquinone FeS Cyt c1 Cyt c Cyt a Cyt a3 1/2 O2

proteins that accept electrons from QH2 or FeS

Ultimately transfers the electrons to oxygen

Mitochondrial Complexes
NAD+ FMN

I
FeS FAD FeS ubiquinone Cyt b

NADH Dehydrogenase

II
Succinate dehydrogenase
ubiquinone

Cytochrome Oxidase

FeS

Cyt c1

Cyt c

Cyt a

Cyt a3 1/2 O2

III
CoQ-cyt c Reductase

IV

Electron Transport chain (ETC)

ETC present in inner mitochondrial membrane as four complexes

Complexes I, III and IV do the pumping of protons

These are transmembrane protein complexes containing quinones, flavin nucleotides, sulfur clusters, hemes, and copper ions. The inner membrane (IMM) is impermeable to ions and polar molecules. Specific transporters shuttle metabolites such as ATP and organic ions.

Oxidative Phosphorylation

Definition:
Oxidative Phosphorylation is the process in which ATP is formed as a result of the transfer of electrons from NADH and FADH2 to O2 by ETC.

Mechanism of Oxidative Phosphorylation ( Chemiosmotic hypothesis and binding change model)

The Electron transport through ETC is coupled with translocation of protons across IMM. This results in the formation of an electrochemical gradient i.e. more H+ on outer side of IMM than inside. The Protons flow back into the mitochondrial matrix and induce a conformational change in the enzyme ATPase which synthesizes ATP from ADP and Pi.

Matrix
H+ + NADH NAD+ + 2H+ 2H+ + O2 H2O

2 eQ

III IV

++
4H+ 4H+

cyt c

2H+

Intermembrane Space

4H+ are pumped per 2e- passing through complex III. The H+/e- ratio is less certain for the other complexes: probably 4H+/2e- for complex I; 2H+/2e- for complex IV.

1.Electrons are transported along the inner mitochondrial membrane, through a series of electron carriers 2.Protons (indicated by + charge) are translocated across the membrane, from the matrix to the intermembrane space 3.Oxygen is the terminal electron acceptor, combining with electrons and H+ ions to produce water 4. As NADH delivers more H+ and electrons into the ETS, the proton gradient increases, with H+ building up outside the inner mitochondrial membrane, and OH- inside the membrane.

OH- ADP3- ATP4ADP3-

INTERMEMBRANE SPACE H+ NADH + H+ H+ eNAD+ H+ 4H+ O2 e-H O 2 complex I FMNH2 eeinner e membrane CoQ complex III e- b cyt e H+ C1 e cyt e a-a3 C eH+ complex IV

OH-

ATP4ADP3ADP3-

OH- ADP3ADP33Pi- ADP3-

ATP4-

3H+

ATP4-

F1

MATRIX

stalk

Fo
Proton gradient/Charge gradient 3H+
Electron transport and Oxidative Phosphorylation

High Energy Compounds

Definition: Certain compounds which on hydrolysis
yield at least 7cal/mol free energy at pH 7.0 are known as high energy compounds.

Classification
Pyrophosphates e.g. ATP Acyl Phosphates e.g.1,3-bisphosphoglycerate Enol Phosphates e.g. Phosphoenolpyruvate Thioesters e.g. Acetyl Co A Phosphagens e.g. Creatine Phosphate

ATP- The Energy Currency of Cell

ATP made up of adenine, ribose and a triphosphate moiety.

ATP- The Energy Currency of Cell-contd ATP is a unique and most important high energy molecule in the body due to presence of two high energy phosphoanhydride bonds. ATP is hydrolyzed to yield energy ATP + H2O ADP +Pi + 7.3 Cal ATP serves as the Energy currency of Cell as it is being constantly utilized and regenerated in ATP-ADP cycle. ATP also acts a phosphate donor to low energy phosphate compounds. ATP serves as a link between Catabolism and Anabolism in biological system.

Storage forms of high Energy Phosphate

Creatine Phosphate (CP) is the reservoir of energy in muscles. CP donates its phosphate to ADP to form ATP by Enzyme Creatine Phosphokinase. When muscles contract (active muscle) energy is derived from breakdown of ATP to ADP and Pi. ATP must be reformed quickly to supply the energy which initially comes from creatine phosphate and subsequently from glycolysis. In resting muscle Creatine is again phosphorylated and energy is stored as CP.

G0 Compound (kJ/mol) ( kcal/mol) Phosphoenolpyruvate 61.9 14.8 Carbamoyl phosphate 51.4 12.3 1,3-Bisphosphoglycerate 49.3 11.8 (to 3-phosphoglycerate) Creatine phosphate 43.1 10.3 ATP ADP + Pi 30.5 7.3 ADP AMP + Pi 27.6 6.6 Pyrophosphate 27.6 6.6 Glucose 1-phosphate 20.9 5.0 Fructose 6-phosphate 15.9 3.8 AMP 14.2 3.4 Glucose 6-phosphate 13.8 3.3 Glycerol 3-phosphate 9.2 2.2

Thank you very much with All the best wishes