Cellular Respiration and ATP Synthesis

Pt 1

Learning Objectives
At the end of this document students should be able to:

 State what is cellular respiration.

 Describe the importance of a mitochondria,
relating its structure to its function and create an
annotated diagram of the mitochondrion.
 Outline the stepwise breakdown of glucose in
cellular respiration.
 Explain the sequence of steps in glycolysis.
 Explain the significance of glycolysis.

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Introduction to Cellular Respiration
- Cellular respiration is the series of metabolic reactions that takes place within a cell in
which the biochemical energy in organic substances is harvested/extracted and then
stored in energy carrying biomolecules. Cellular respiration can be aerobic or anaerobic.

- Aerobic Respiration: the oxidation of organic compounds with oxygen to release energy.

- Cellular Respiration releases heat (unusable energy) and free electrons. The free electrons
serve as a source of energy for producing ATP. ATP then serves as the energy source for
most of the body’s endergonic reactions. The process of cellular respiration is a series of
coordinated enzyme-catalyzed reactions that capture energy from biological
macromolecules.

- When O2 is present in the cell, aerobic respiration takes place. Aerobic respiration is the
process by which organisms use oxygen to turn fuel, such as fats and sugars, into
chemical energy.

o 6O2 + C6H12O6 → 6CO2 + 6H2O + Free Energy +Heat Energy

- Without O2 present in the cell, anaerobic respiration takes place. Anaerobic

respiration is catabolism of organic compounds using electron acceptors other than
molecular oxygen (O2).

RESPIRATION AND REDOX REACTIONS

Aerobic Respiration: Role of NAD and FAD

 Coenzymes NAD (Nicotinamide Adenine Dinucleotide) and FAD (Flavine Adenine
Dinucleotide) play a critical role in aerobic respiration.
 When hydrogen atoms become available at different points during respiration NAD and
FAD accept these hydrogen atoms.
o A hydrogen atom consists of a hydrogen ion and an electron.
 When the coenzymes gain a hydrogen they are ‘reduced’
o NADH +H/ FADH2
o OIL RIG: Oxidation Is Loss, Reduction Is Gain
 When the hydrogen atoms are removed the coenzymes are ‘oxidised’

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 o NAD+/FAD
 They are responsible for accepting "high energy" electrons from the different stages of
respiration and carrying them ultimately to the mitochondria where they are used to
synthesize ATP molecules.
 Hydrogen ions and electrons are important as they play a role in the synthesis of ATP

The reduction and oxidation of NAD and FAD.

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MITOCHONDRIA - ATP SYNTHESIS AND CELLULAR
RESPIRATION
Structure & Function of the Mitochondria

 Mitochondria are rod-shaped organelles 0.5 - 1.0 µm in diameter

 They are the site of aerobic respiration in eukaryotic cells
 The function of mitochondria is to synthesize ATP
 Synthesis of ATP in the mitochondria occurs during the last stage of respiration called
oxidative phosphorylation
o This relies on membrane proteins that make up the ‘electron transport chain’ and
the ATP synthase enzyme.

The structure of a mitochondrion

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Mitochondria structure

1. Mitochondria have two phospholipid membranes:

o The outer membrane is:
 Smooth
 Permeable to several small molecules (pyruvate, oxygen, CO2, ATP, ADP
but not glucose)
o The inner membrane is:
 Folded (cristae) to give a large surface area
 Less permeable (permeable to pyruvate, oxygen, CO2, ATP, ADP but
not glucose but especially not permeable to hydrogen ions)
 The site of the electron transport chain (used in oxidative
phosphorylation)
 Location of ATP synthase (used in oxidative phosphorylation)

2. The intermembrane space:

o Has a low pH due to the high concentration of protons
o The concentration gradient across the inner membrane is formed during oxidative
phosphorylation and is essential for ATP synthesis

3. The matrix:
o Is an aqueous solution within the inner membranes of the mitochondrion.
o Contains ribosomes, enzymes, and circular mitochondrial DNA necessary for
mitochondria to function.
o This is where the link reaction, and the Krebs cycle occur.

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 Electron micrograph of mitochondria

The structure of the mitochondrion is adapted to the function it performs:

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  Outer membrane – the outer membrane contains transport proteins that enable the
shuttling of pyruvate from the cytosol.

 Inner membrane – contains the electron transport chain and ATP synthase (used for
oxidative phosphorylation).

 Cristae – the inner membrane is arranged into folds so that is has a large surface which
enables the membrane to hold many electron transport chain proteins and ATP synthase
enzymes.

 Intermembrane space – small space between membranes maximizes hydrogen gradient

upon proton accumulation.

 Matrix – central cavity that contains appropriate enzymes and a suitable pH for the Krebs
cycle to occur.

THE FOUR STAGES OF AEROBIC RESPIRATION

 The process of aerobic respiration using glucose can be split into four stages. Each stage
occurs at a particular location in a eukaryotic cell.

o Glycolysis takes place in the cell cytoplasm.

o The Link reaction takes place in the matrix of the mitochondria.
o The Krebs cycle takes place in the matrix of the mitochondria.
o Oxidative phosphorylation occurs at the inner membrane of the mitochondria.

Four Stages of Respiration

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Exam Tip
It’s important to know the exact locations of each stage. It is not enough to say the Krebs cycle
takes place in the mitochondria, you need to say it takes place in the matrix of the mitochondria.

Aerobic Respiration: Glycolysis

 Glycolysis is the first stage of respiration. Glucose is the main respiratory substrate used
by cells for this process.
 Glycolysis is a catabolic chemical process that involves complex molecules being broken
down into smaller one to release energy.
o The energy made during glycolysis involves phosphorylation (attachment of a
phosphate group to a molecule) to form ATP. ATP can be formed by different
types of phosphorylation;
 Substrate-linked phosphorylation – this is where a phosphate is transferred
from a substrate directly to ADP. This is the production of ATP or GTP

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 supported by the energy released from another high-energy bond that leads
to phosphorylation of ADP or GDP to ATP or GTP.

 Chemiosmotic Phosphorylation - The production of ATP using the process

of chemiosmosis - the movement of ions across a membrane in an
electrochemical gradient.
 Photophosphorylation – this is where the energy of sunlight is used to
form ATP.

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  Glycolysis takes place in two phases in the cytoplasm of the cell and involves:
o An energy investment phase – step 1 - 5
o An energy releasing phase – steps 6 - 10
 It results in the production of:
o 2 Pyruvate (3C) molecules
o Net gain 2 ATP
o 2 reduced NAD
Enzymes used in glycolysis:
- Kinases - an enzyme that catalyzes the transfer of a phosphate group from
ATP to a specified molecule.
- Dehydrogenases - an enzyme that catalyzes the removal of hydrogen atoms
from a particular molecule. Dehydrogenases are mainly responsible for
oxidation of its substrates.
- Isomerases - an enzyme that catalyzes the conversion of a specified compound
to an isomer.
- Phosphatases are enzymes that specifically remove phosphate groups from
their substrates.

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Steps of glycolysis
 Step 1: Phosphorylation
o Glucose (6C) is phosphorylated by ATP to form glucose 6-phosphate (6C). This
is done to make the glucose more reactive and to lower the activation energy of
the reaction. This phosphorylation also traps the glucose in the cell and prevents it
from being transported out of the cell.

 Step 2: Isomerization of Glucose-6-phosphate

o Glucose 6-phosphate is converted to it isomer frustose-6-phospate.
 Why is the isomerization of glucose-6-phosphate to fructose-6-phosphate
important?
 This is done so that carbon 6 can be exposed to attach the second
phosphate group from ATP.
 It is also needed because fructose can be cleaved to yield two 3 carbon
compounds rather than glucose.

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  Step 3: Phosphorylation of Frustose-6-phosphate
o Another phosphate group gets transferred from ATP to fructose-6-phosphate to
produce fructose-1,6-bisphosphate. This is done, so that it can now interact with
different enzymes in a way it couldn't when it was just a monophosphate.

o The advantages of converting glucose-6-phosphate to fructose-6-phosphate, and

then phosphorylating it to fructose-1,6-biphosphate is that fructose-1,6-
biphosphate can be easily cleaved into two symmetrical phosphorylated
interconvertible three-carbon compounds.

 Step 4: Lysis: Cleavage of Fructose 1,6 bisphosphate

o Fructose bisphosphate (6C) splits into two molecules of triose phosphate (3C)
dihydroxyacetone phosphate (DHAP) and glyceraldehyde 3-phosphate. These two
three-carbon molecules are isomers of each other. Out of the two, only
glyceraldehyde-3-phosphate can directly continue through the next steps of
glycolysis.

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 o Fructose bisphosphate → 2 Triose phosphate

 Step 5: Isomerization of DHAP

o As DHAP cannot directly take part in the consecutive steps of the process, it gets
converted into its isomer glyceraldehyde-3-phosphate. Hence two molecules of
glyceraldehyde 3-phosphate are formed from one molecule of glucose.

 Step 6: Oxidation and phosphorylation

o Next, the enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH) adds
phosphate from the cytosol to the glyceraldehyde 3-phosphate to form 1,3-
bisphosphoglycerate. ATP is not required; the source of the phosphate group is a
simple phosphate ion.
o The same enzyme also dehydrogenates glyceraldehyde 3-phosphate by removing
two of its hydrogens and transferring it to the oxidizing agent NAD+ to form
NADH⁺. The two molecules of glyceraldehyde 3-phosphate both undergo this
process of phosphorylation and dehydrogenation.

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  Step 7: Dephosphorylation
o The enzyme phosphoglycerate kinase transfers the high-energy phosphoryl group
from the carboxyl group of 1,3-bisphosphoglycerate to ADP, forming ATP and 3-
phosphoglycerate.
o This step generates ATP via substrate linked phosphorylation.
o As there are two molecules of 1,3-bisphosphoglycerate, the reaction yields two 3
PGA, and 2 ATPs.

 Step 8: Intermolecular shift of phosphate group

o Phosphoglycerate mutase shifts the phosphate group from 3rd to 2nd carbon atom,
converting 3-phospho glycerate to 2-phospho glycerate.

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 o This places the phosphate in a favorable position for its removal to form ATP in
later reactions.

 Step 9: Dehydration of 2-Phosphoglycerate

o An enzyme removes a molecule of water from 2-phosphoglycerate converting 2-
PG into phosphophenol pyruvate (PEP).

 Step 10: Conversion of PEP to pyruvate

o As PEP is an unstable molecule, it loses one of its phosphate group in this step.
PEP is converted to pyruvate by the removal of its phosphate group by pyruvate
kinase. This step yields 2 molecules of pyruvate and 2 ATPs in the end. Two ATP
molecules are used in glycolysis, and at the four ATP are produced.

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  Pyruvate is considered as the end product of glycolysis.
2 Triose phosphate → 2 Pyruvate

Summary of Glycolysis

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Function & Advantage of Glycolysis

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  The main advantage of glycolysis is that it produces energy very fast, without requiring
oxygen.
 It breaks down glucose to form high-energy molecules ATP and NADH, which fuel the
cellular activities.
 Pyruvate, the end product of this process, helps to continue the chain reactions of cellular
respiration. In aerobic respiration it enters the citric acid cycle, whereas in anaerobic
respiration it takes part in fermentation.
 The intermediate compounds formed in this process can be used in various other cellular
processes.
Summary of Glycolysis

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