CARBOHYDRA TES

Carbohydrates
Bioenergetics and Metabolism
Lecture 5

Lecture - 5

Metabolic Pathways:
Definition Anabolism Catabolism & Catabolic stages

Regulation of metabolism:
Signals from within the cell Communication between cells Membrane receptor Intracellular messenger systems

Metabolism

Is the sum of all chemical changes occurring in a cell, a tissue, or the body.

Metabolic pathways:
Definition:

It is formed of a sequence of enzymatic reactions where the product of an enzymatic reaction becomes the substrate for the next reaction; the successive products of the reactions known as metabolites or metabolic intermediates.

Metabolic pathways Metabolism is classified into: Anabolism and Catabolism

I- Anabolism

It includes all the biosynthetic pathways which are concerned with synthesis of complex end products from simple precursors, for example, synthesis of glycogen, proteins and lipids from simple molecules. Anabolic reactions require energy which is supplied mainly by adenosine triphosphate (ATP).

II- Catabolism It includes all degradative processes whereby complex molecules, such as proteins, polysachharides, and lipids, are broken into a few simple molecules , for example, CO2, NH3 and water. Catabolism is classified into three main stages:

Stage I: Hydrolysis of complex molecules: Complex molecules are broken down into their component building blocks. For example, proteins are degraded to amino acids, polysaccharides to monosaccharides, and triacylglycerols to free fatty acids and glycerol.

Stage II: Conversion of building blocks to simple intermediates: The diverse building blocks are further degraded to acetyl CoA and a few other, simple molecules. Some energy is captured as ATP.

Stage III: Oxidation of acetyl CoA: The tricarboxylic acid (TCA) cycle is the final common pathway in the oxidation of fuel molecules such as acetyl CoA. Large amounts of ATP are generated as electrons flow from NADH and FADH2 to oxygen via oxidative phosphorylation.

Carbohydrates
Stage One
Monosaccharides

Proteins
Stage One
Amino Acids

Lipids
Stage One
Glycerol + Fatty Acids

Stage Two
Acetyl-CoA OR Intermediates of Citric Acid Cycle

Stage Three
Krebs Cycle
Reduced Coenzyme

ETC

ATP + H2O
2 CO2 + ATP

Bioenergetics

LOW AND HIGH ENERGY BONDS

When ATP is hydrolyzed to ADP + Pi, the energy released is about 7.3 Kcal /mole (7300 cal/mole). Chemical bonds are mainly two types as follows: I- Low Energy Bonds II- High Energy Bonds

I- Low Energy Bonds These are bonds that on hydrolysis produce an amount of free energy less than 7 Kcal/mole and include most of chemical bonds, for example: 1- Phosphate ester bonds e.g. glucose -6- P or glucose -1- P. 2- Glycosidic bonds in carbohydrates. 3- Peptide bonds in proteins.

II- High Energy Bonds These are bonds which on hydrolysis produce an amount of free energy more than 7 (7.3 - 14.8) Kcal/mole. They include the following:
High Energy Phosphate Bonds :
O

~P
OH

OH

1- Enol-Phosphate: ex: 2-phospho enol pyruvate COOH C CH2 O~ P

2- Carboxylic ~ Phosphate bonds: ex: a) alpha-1,3-Bisphosphoglycerate O C O

~
O

H C OH CH2 P

ATP

ATP is a high energy compound. It consists of adenosine (adenine + ribose) to which three phosphate groups are attached. If one phosphate is removed, adenosine diphosphate (ADP) is produced; if two phosphates are removed, adenosine monophosphate (AMP) results. The standard free energy of hydrolysis of ATP is approximately -7300 cal/mole for each of the two terminal phosphate groups.

Adenosine Triphosphate (ATP)

ATP-ADP Cycle

Oxidative Phosphorylation

~
ATP

ADP

Creatine

Muscles

~
Muscle Contraction

P Creatine ~ Phosphorylation of compounds P

Active transport Anabolism

Nerve Impulses

Mechanism Of Collection Of Energy:

Free energy liberated during the degradation of foodstuffs is collected in the form of high energy phosphate bonds at 2 levels in metabolism. At the substrate level and At the respiratory chain level.

A) Substrate Level Phosphorylation:

Pyruvate Kinase
Phosphoenolpyruvate + ADP

Enolpyruvate +

ATP

Succinate thiokinase
Succinyl-CoA + ADP + Pi Succinic Acid + ATP

B) Oxidative Phosphorylation:

CARBOHYDRA TES

Electron Transport Chain (ETC)

Lecture - 6
Electron Transport Chain:
Overview Components of ETC Chemiosmotic Hypothesis Synthesis of ATP (ATP Synthase) Control of oxidation in ETC Inhibitors of ETC Uncouplers

Energy-rich compounds , such as carbohydrates, fats and proteins are metabolized by a series of oxidation reactions yielding CO2 and water.
The metabolic intermediates of these reactions donate electrons to specific coenzymes nicotineamide adenine dinucleotide (NAD+) and flavin adenine dinucleotide (FAD) to form the energy-rich reduced coenzymes, NADH and FADH2. These reduced coenzymes can, in turn, each donate a pair of electrons to a specialized set of electron carriers, collectively called the electron transport chain.

An electrons are passed down the electron transport chain, they lose much of their free energy.

Part of this energy can be captured and stored by the production of ATP from ADP and inorganic phosphate (Pi).
This process is called oxidative phosphorylation.

Active acetate

Kreb's Cycle

Reduced Coenzymes + 2CO2

O ETC Oxidized Coenzymes + H2O

Energy ATP
Phosphorylation

ADP + Pi

ELECTRON TRANSPORT CHAIN (ETC) RESPIRATORY CHAIN

ETC is formed of a series of electron carriers, which catalyze the transfer of electrons from reduced coenzymes to oxygen to form H20. Part of the energy released is utilized for synthesis of high-energy phosphate bonds (i.e. conversion of ADP + Pi to ATP).

Components of ETC The components of the ETC are located in the inner mitochondrial membrane and is the final common pathway by which electrons derived from different fuels of the body flow to oxygen. Electron transport and ATP synthesis by oxidative phosphorylation proceed continuously in all tissues that contain mitochondria. Note: - The inner mitochondrial membrane is impermeable to most small ions, including H+, Na+, and K+, small molecules such as ATP, ADP, pyruvate, and other metabolites important to mitochondrial function. - Specialized carriers or transport systems are required to move ions or molecules across this membrane.

Components of ETC It is formed of four complexes and 2 mobile electron carrier (coenzymes Q and cytochrome c).

Electron transport chain

Organization of ETC The inner mitochondrial membrane contains five enzyme complexes, called I, II, II, IV, and V. Complexes I to IV each contain part of the ETC, whereas complex V catalyzes ATP synthesis. Each complex accepts or donates electrons to relatively mobile electron carriers, such as coenzyme Q and cytochrome c. Each carrier in the ETC can receive electrons from an electron donor, and can consequently donate electrons to the next carrier in the chain. The electrons combine with oxygen and protons to form water.

Structure of a mitochondrion showing the electron transport chain and ATP synthesizing structures on the inner membrane

Oxidative Phosphorylation

The transfer of electrons down the electron transport chain is energetically favored because NADH is a strong electron donor and molecular oxygen is an avid electron acceptor. However, the flow of electrons from NADH to oxygen does not directly result in ATP synthesis.

Chemiosmotic hypothesis of ATP synthesis

The chemiosmotic hypothesis explains how the free energy generated by the transport of electrons by the electron transport chain is used to produce ATP from ADP + Pi. Electron transport is coupled to the phosphorylation of ADP by the transport of protons (H+) across the inner mitochondrial membrane from the matrix to the intermembrane space.

Chemiosmotic hypothesis
1. Proton pump: It is proved that the energy of electron transport through the ETC is utilized by complex I, III and IV that act as proton pumps, for transfer of protons from the matrix to the inter-membrane space. This process creates, across the inner mitohondrial membrane, an electerical gradient (with more positive charges on the outside of the membrane than on the inside) and a pH gradient (the outside of the membrane is at a lower pH than the inside). The energy generated by this proton gradient is sufficient to drive ATP synthesis.

Chemiosmotic hypothesis
2. ATP synthase (complex V): This enzyme complex synthesizes ATP, using the energy of the proton gradient generated by the ETC. After protons have been transferred from the matrix to the intermembrane space, they reenter matrix by passing through a channel in the ATP synthase complex, resulting in the synthesis of ATP from ADP + Pi and, at the same time, dissipating the pH and electerical gradient.

Chemiosmotic Hypothesis

Chemiosmotic hypothesis
Oligomycin: This drug binds to ATP synthase, closing the H+ channel, and preventing reentry of protons into the mitochondrial matrix. Because the pH and electerical gradients cannot be dissipated, electron transport stops. As electron transport and phosphorylation are tightly coupled, inhibition of phosphorylation inhibits oxidation.

Control of Oxidation in ETC

A supply of ADP is necessary for ATP synthesis; a low concentration of ADP will result in decreased production of ATP. Since electron transport and ATP synthesis are tightly coupled, electron transport and thus oxidation of NADH and FADH2 will also be inhibited.

Inhibitors of ETC

These compounds prevent the passage of electrons by binding to a component of the respiratory chain, blocking the oxidation-reduction reaction. Therefore, all electron carriers before the block are fully reduced and those after the block are oxidized. Because electron transport and oxidative phosphorylation are tightly coupled, ATP synthesis is also inhibited.

Site-Specific inhibitors of electron transport

Uncouplers of ETC

They act to dissociate oxidation in the ETC from phosphorylation (ATP synthesis) and energy released as heat. Electron transport and phosphorylation can be uncoupled by compounds that increase the permeability of the inner mitochondrial membrane to protons e.g. 2,4-dinitrophenol.

Oxidation Of Extra-Mitochondrial NADH

NADH is produced in the cytosol during oxidation of glucose by glycolysis. NADH cannot pass through the mitochondrial membrane to reach the ETC for its oxidation. Instead, two shuttles can function in the transfer of hydrogen (Reduced Equivalent) from NADH in the cytosol to mitochondria: Glycerophosphate Shuttle.

Malate - Aspartate Shuttle.

Glycerophosphate Shuttle

Malate-Aspartate Shuttle