Introduction to Metabolism
&
Biological Oxidation
By
Dr. Salwa Abo Elkhair
Professor of Medical Biochemistry & Molecular Biology
[email protected]
Outline
Introduction to Metabolism
ATP as an energy carrier
Biological Oxidation
Electron Transport Chain
Oxidative Phosphorylation
Metabolism
Metabolism
• Energy
– A central theme of Biochemistry
– Can be produced by oxidation of foods: glucose, FA, aa
• Energy is essential to:
– Grow, maintain structure & function of the cell
•Metabolism is the sum total of all the reactions
that take place in a living cell.
Metabolism = Catabolism + Anabolism
Catabolism - larger molecules are broken down into smaller ones in a
process that usually releases energy
Anabolism - larger molecules are made from small ones in a process the
usually requires energy
Metabolism
• ATP is used to shuttle chemical energy
from catabolism to anabolism.
Metabolism
Metabolic Sources of Energy
So, Energy is produced by oxidation of food stuffs with the production of the
end products CO2 and H2O.
• This energy is required to maintain structure and function of the living cells.
Applied Aspects
Indigestion & Malabsorption: can cause deficiencies
of all nutrients (primary metabolism) leading to
weight loss & fatigue.
Cancer & weight loss: result partly from loss of
appetite, change of eating habits and the way body
processes & absorbs food (primary metabolism).
Also, cancer cells put the body in a catabolic state of
metabolism (Cancer cells demand more energy) &
makes body breaking down muscle tissue and fat
(Cancer cachexia).
ATP
ATP: an energy carrier
ATP is a high-energy phosphate compound.
The energy currency of cell
Structure: adenine + ribose + 3 inorganic
phosphates
Contain 2 high energy phosphate bond.
Function of ATP
• Biological Importance of ATP: Source of energy for:
1. Biosynthetic reactions.
2. Biosynthesis of cAMP.
3. Muscle contraction.
4. Nerve conduction.
5. Active absorption and secretion.
6. Active transport across biological membranes.
7. Activation of monosaccharides, FA and AA.
8. Formation of creatine phosphate: energy store in muscle.
ATP Production
How do cells make ATP
By PHOSPHORYLATION... adding a phosphate to ADP
ADP + P ------> ATP
2 mechanisms of phosphorylation:
A. Substrate level phosphorylation.
B. Oxidative phosphorylation.
ATP Production How do cells make ATP
(1) Substrate level phosphorylation
ATP can be formed (during metabolic pathways e.g. glycolysis ,
Krebs cycle) by transferring the high energy phosphate group
from substrates directly to ADP.
A substrate molecule ( X-p ) donates its high energy P to ADP
making ATP
How is ATP produced
(2) Oxidative phosphorylation
The electrons produced by oxidation of foodstuffs are transferred to react
finally with oxygen and the produced energy is utilized for ATP synthesis.
i.e it is the process in which ATP is formed as a result
of the transfer of electrons from NADH or FADH2 to O2
by a series of electron carriers.
Biological Oxidation
Biological oxidation
• Biological oxidation is that oxidation which occurs in
biological systems to produce energy.
Oxidation can occur by:
1. Addition of oxygen (less common)
2. Removal of hydrogen (common)
3. Removal of electrons (most common)
Oxidation-Reduction Reactions
•Redox: reduction-oxidation reaction
•Oxidation is loss of electrons
•Reduction is gain of electrons
• Redox potential is a measure of the affinity of substance
to acquire electrons and thereby be reduced.
Hydrogen and Electron carriers
• When food molecule oxidized – electrons and hydrogen are removed.
• By:
1- NAD+ (nicotinamide adenine dinucleotide)
2- FAD (flavin adenine dinucleotide)
• Reduced form is : NADH+H+ and FADH2
* NADH+H+ and FADH2
- transfer e- to O2 in the mitochondria by means of Electron
Transport Chain > ATP generated in this process
Electron Transport Chain
(ETC)
Electron Transport Chain (ETC)
Respiratory
Electron TransportChain,
Chain Redox Chain
• It is a chain of catalysts that Collects reducing equivalents (hydrogen
atoms and electrons) from substrates transferring stepwise to be
oxidized in a final reaction with oxygen (electrons combine with O2 and
protons) to form water and energy (Oxidative Phosphorylation).
• Associated with cell breath, also, called respiratory chain.
• Significance of Electron transfer chain
– Electron transport and oxidative phosphorylation
re-oxidize NADH and FADH2 and trap the energy
released as ATP.
– It represents the final stage in the oxidation of
carbohydrates, fats, and amino acids.
• Site: Located in the inner membrane of mitochondria.
Electron Transport Chain
(ETC)
Components of the electron transport chain
1. Hydrogen and electron carriers.
FAD
NAD→ FMN→ Co Q→ Cyt b→ c1 → c → a → a3
2. Four membrane-bound enzyme complexes.
• All are embedded in inner mitochondrial membrane.
Hydrogen and electron carriers of ETC
FMN
Cyt a,a3
Cyt b,c1
Enzyme Complexes of ETC
Complex I: NADH dehydrogenase.
Complex II: Succinate dehydrogenase.
Complex III: Ubiquinol dehydrogenase.
Complex IV: Cytochrome oxidase.
Complex V: ATP synthase.
Enzyme Complexes of ETC
FMN
Cyt a,a3
Cyt b,c1
• Complex I: NADH dehydrogenase
• It is a flavoprotein that contains FMN as well as FeS protein as coenzymes.
• It transfers hydrogen atoms from NADH+H+ to CoQ.
• It releases 4H+ to interspace of inner and outer membrane.
Enzyme Complexes of ETC
FMN Cyt a,a3
Cyt b,c1
• Complex II: Succinate dehydrogenase
• It is a flavoprotein that contains FAD as well as FeS protein as coenzymes.
• It transfers hydrogen atoms from succinate to CoQ.
• It does not release H+ to the interspace.
Enzyme Complexes of ETC
FMN
Cyt a,a3
Cyt b,c1
• Complex III: Ubiquinol dehydrogenase
• It transfers electrons from CoQ to cytochrome c using cyt b and cyt c1 as coenzymes.
• Every two electrons' transferring lead 4 H+ pumped to the inter-membrance space.
Enzyme Complexes of ETC
FMN Cyt a,a3
Cyt b,c1
• Complex IV: Cytochrome oxidase
• It needs cyt a and cyt a3 as coenzymes.
• It transfers electrons from cytochrome c to oxygen, converting O2 to H20.
• Every two electrons' transferred lead 2 H+ pumped to the intermembrance
space.
• The only electron carrier can directly react with molecular oxygen.
Enzyme Complexes of ETC
FMN
Cyt a,a3
Cyt b,c1
Complex V: ATP synthase.
COMPONENTS OF ETC
6 important Components:
• complex I: NADH dehydrogenase
• complex II: Succinate dehydrogenase
• Mobile carriers: Ubiquinone (CoQ)
• complex III: ubiquinol dehydrogenase complex
• Mobile carriers: Cytochrome c (Cyt c)
• complex IV: Cytochrome c oxidase
NADH → Complex I → Q → Complex III → cyt c → Complex IV → O2
Complex II
• In addition to:
• Complex V = ATP Synthase
Summary: Two respiratory chain
Succinic acid oxidation
Succinic acid respiratory chain
FAD complexesⅡ
NADH oxidation (Fe-S)
respiratory chain
NADH FMN CoQ Cyt b→Cyt c1 Cyt c Cyt aa3 1/2O2
(Fe-S)
Complexes Ⅲ Complexes Ⅳ
complexesⅠ
Intermembrane
space
2H+
4H+ 4H+ Cyt c
+ + + + + + + + + +
I Q F
0
Ⅱ Ⅳ
- - - Ⅲ
- - - - - -
NADH+H+ fumarate H2 O
1/2O2+2H+ F1
NAD+ Succinate
Matrix
ADP+Pi ATP
H+
Applied Aspects
Iron deficiency in anemia affects the cytochromes
and FeS protein as well.
Fatigue in this case results partly from the lack of
electron transport for ATP production.
Oxidative Phosphorylation
Oxidative Phosphorylation
• It means coupling of the electron transport in respiratory chain with
phosphorylation of ADP to form ATP (by Complex V = ATP Synthase).
• It is a process by which the energy of biological oxidation is ultimately
converted to the chemical energy of ATP.
• There are 3 sites ( complex I, III, IV).
Mechanism of oxidative phosphorylation
Chemiosmotic theory (Mitchell Hypothesis).
It suggest that the transfer of electrons through the electrons
transport chain causes protons to be translocated (pumped out)
from the mitochondrial matrix to the intermembrane space at the
three sites of ATP production (i.e. it acts as a proton pump (H+) ).
resulting in an electrochemical potential difference (proton
gradient) across the inner mitochondrial membrane.
Flow of H+ back is used to drive the conversion of ADP+Pi to ATP
+
+ __ + +
_ _
+ _ _ + __ +
+ _ + __ +
+ _+ _ _
+ _ _ _ _ +
+ _ + __ _
_ +_ +
+ + + __ _ _
+ _ _ +
+ _ + _ _ +
+ + _
+ +
+ + +
Starting via NAD, 3 ATP are formed for each substrate molecule oxidized.
Starting via FAD, 2 ATP are formed for each substrate molecule oxidized.
P/O Ratio
• The relationship between ATP synthesis and O2 consumption.
• It is a measure to the efficiency of oxidative phosphorylation.
the number of molecules of ATP made
P/O ratio = -----------------------------------------------
the number of O2 atoms consumed
• When NADH is used, P/O = 3/1
• When FAD is used, P/O = 2/1
succinate The sites of coupled oxidative
phosphorylation
Regulation of Oxidative Phosphorylation
Respiratory control is based on a tight coupling between electron flow
in the respiratory chain and ATP synthesis.
The rate of ATP synthesis is dependent on the availability of
substrates.
The possible factors being
NADH
Oxygen
ADP
phosphate
Uncouplers
They are substances that dissociate oxidation from phosphorylation leading to loss
of the resulting energy as heat.
There is normal oxygen consumption without ATP generation and P/O ratio
becomes zero.
They increase membrane permeability to protons which leads to transport of
protons into the mitochondrial matrix reducing the electrochemical potential
difference, so, ATP will not be generated.
Example of the uncouplers include:
1. Calcium injection.
Loss of energy as heat
2. Dicoumarol, used as anticoagulant.
3. Thyroid hormones.
Hyperthermia & Heat Intolerance
4. Progesterone.
Applied Aspects
In auto-immune induced hyperthyroidism (Graves’
disease), increased thyroid hormones lead to loss
of the energy as heat. So, patient complains of
hyperthermia & heat intolerance.
Hyperthermia & heat intolerance in Pregnant female
result partly from the increased progesterone
(hormone of pregnancy).
Applied Aspects
In high doses, the drug aspirin (as well as other
salicylates) uncouples oxidative phosphorylation. This
explains the fever that accompanies toxic overdoses of
these drugs.
Points
• Function of ATP
• How is ATP produced
– Substrate level phosphorylation
– Oxidative phosphorylation
• Electron transfer chain (or Respiratory chain)
– Definition, Significance, Site, Components
– Two respiratory chain
• NADH oxidation respiratory chain
• Succinic acid oxidation respiratory chain
• Oxidative Phosphorylation
– P/O Ratio
– 3/1 ATP per NADH; 2/1 ATP per FADH2
• Uncouplers.
