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CHAPTER 8

RESPIRATION
8.1 ATP (Adenosine Triphosphate)

Immediate energy source that drives


most cellular work.
Cells do this work by energy
coupling - uses an exergonic
process to drive an endergonic one.
Endergonic Reaction - ends with net
gain in energy
Products have more energy than
reactants, e.g., photosynthesis
Exergonic Reaction - ends with net
loss in energy
Reactants have more energy than
products, e.g., cellular respiration
Structure and hydrolysis of ATP

ATP = Nucleotide with unstable


phosphate bonds that cell hydrolyzes
for energy to drive endergonic
reactions.
Consists of adenine, ribose, & chain
of three phosphate groups.
Unstable bonds between phosphate
groups can be hydrolyzed in an exergonic
reaction.
When terminal phosphate bond is
hydrolyzed, a phosphate group is
removed producing ADP (adenosine
diphosphate).
ATP + H2O → ADP + Pi
Under standard lab conditions, reaction
releases -31 kJ/mol (-7.3 kcal/mol).
In living cell, reaction releases -55
kJ/mol (-13 kcal/mol) - 77% more than
under standard conditions.
Terminal phosphate bonds of ATP
are unstable, so:
– Products of hydrolysis reaction are
more stable than reactants.
– Hydrolysis of phosphate bonds is
thus exergonic as system shifts to a
more stable state.
How ATP performs work
Exergonic hydrolysis of ATP coupled
with endergonic processes - phosphate
group transferred to another molecule.
– Phosphate transfer enzymatically
controlled.
– Molecule receiving phosphate
(phosphorylated/activated
intermediate) becomes more reactive.
The regeneration of ATP
ATP continually regenerated by cell.
 Rapid process: 107 molecules used and
regenerated/sec/cell).
 Reaction: endergonic.

ADP + Pi → ATP
∆ G = + 31 kJ/mol (+7.3 kcal/mol)
Energy to drive endergonic regeneration
of ATP comes from exergonic process of
cellular respiration.
8.2 Aerobic Respiration

Preview of cellular respiration


(See Figure 9.6, Campbell, page 164)

Stages of respiration:
i. Glycolysis
ii. Citric acid cycle
iii. Electron transport chain (ETC)
iv. Oxidative phosphorylation.
Glycolysis - in cytoplasm.
Glucose broken down into two
molecules of pyruvate.
Citric acid cycle - in mitochondrial
matrix.
Completes breakdown of glucose
by oxidizing a derivative of pyruvate
to CO2.
Several steps in glycolysis and citric
acid cycle are redox reactions -
dehydrogenase enzymes transfer
electrons from substrates to NAD+,
forming NADH.
NADH passes electrons to ETC.
Electrons then move from molecule to
molecule until they combine with
molecular O2 and H+ to form water.
As they passed along chain, energy
carried by electrons is used to
synthesize ATP in mitochondrion via
oxidative phosphorylation.
Inner membrane of mitochondrion is
site of electron transport chain (ETC)
and chemiosmosis
ETC + Chemiosmosis = Oxidative
Phosphorylation.
Produces almost 90% of ATP
generated by respiration.
Some ATP formed directly during
glycolysis and citric acid cycle by
substrate-level phosphorylation.
– Enzyme transfers phosphate group from
an organic substrate to ADP, forming
ATP.

Enzyme
Enzyme

ADP

P
Substrate + ATP

Product
For each molecule of glucose
degraded to CO2 & H2O, cell makes
up to 38 ATP
Each ATP ≈ 7.3 kcal/mol of free
energy.
8.2.1 Glycolysis

Energy investment phase

Glucose

2 ADP + 2 P 2 ATP used


Glycolysis Citric
acid Oxidative
cycle phosphorylation

Energy payoff phase


ATP ATP ATP 4 ADP + 4 P 4 ATP formed

2 NAD+ + 4 e– + 4 H+ 2 NADH + 2 H+

2 Pyruvate + 2 H2O

Net
Glucose 2 Pyruvate + 2 H2O
4 ATP formed – 2 ATP used 2 ATP
2 NAD+ + 4 e– + 4 H+ 2 NADH + 2 H+
Glucose split into two 3C sugars.
Sugars oxidized and rearranged to form
two molecules of pyruvate, ionized form
of pyruvic acid.
Two phases of glycolysis:
1. Energy investment phase
 Cell invests ATP to provide activation
energy by phosphorylating glucose.
 Requires 2 ATP per glucose.
2. Energy payoff phase
 ATP produced by substrate-level
phosphorylation.
 NAD+ reduced to NADH by electrons
released by oxidation of glucose.
Net yield from glycolysis = 2 ATP and 2
NADH per glucose.
 No CO produced during glycolysis.
2

Glycolysis occurs in presence/absence of


O2.
Glycolysis Citric Oxidation
acid
cycle phosphorylation

The Glycolytic Pathway ATP ATP ATP


Glucose

ATP

Hexokinase

ADP

Glucose-6-phosphate

Phosphoglucoisomerase

Fructose-6-phosphate

ATP

Phosphofructokinase

ADP

Fructose-
1, 6-bisphosphate

Aldolase

Isomerase

Dihydroxyacetone Glyceraldehyde-
phosphate 3-phosphate
2 NAD+
Triose phosphate
dehydrogenase

2 NADH
+ 2 H+

1, 3-Bisphosphoglycerate

2 ADP

Phosphoglycerokinase
2 ATP

3-Phosphoglycerate

Phosphoglyceromutase

2-Phosphoglycerate

Enolase
2 H2O

Phosphoenolpyruvate
2 ADP

Pyruvate kinase
2 ATP

Pyruvate
Overall reaction showing all reactants
and products resulting from
glycolysis:

Glucose + 2ATP + 2Pi + 4ADP + 2NAD+

2 Pyruvate + 2ADP + 4ATP + 2NADH +


2H+ + 2H2O
Equation showing net reaction of
glycolytic pathway.

Glucose + 2Pi + 2ADP + 2NAD+

2 Pyruvate + 2ATP + 2NADH + 2H+ +


2H2O
8.2.2 Pyruvate oxidation
> ¾ of original energy in glucose still
present in the 2 molecules of pyruvate.
If O2 is present, pyruvate enters
mitochondrion where oxidation to CO2 is
completed.
After pyruvate enters mitochondrion via
active transport, it is converted to acetyl
coenzyme A (acetyl CoA).
Involve multienzyme complex that
catalyzes three reactions:
CYTOSOL MITOCHONDRION

NAD+ NADH + H+

Acetyl Co A
Pyruvate CO2 Coenzyme A

Transport protein
1) Carboxyl group removed as CO2.
2) Remaining 2C fragment oxidized to
acetate. An enzyme transfers 2
electrons to NAD+ to form NADH.
3) Acetate combines with coenzyme A,
forming the very reactive acetyl CoA.

Acetyl CoA enters citric acid cycle for


further oxidation.
Formation of Acetyl CoA
O O
C C
O C C C Pyruvat C C C O
es
NA NA
D+ D+
NA NA
DH DH
C C C C C C
CoA CoA

Acetyl
CoA C C
CoA
C C CoA
Glycolysis Citric Oxidation
acid
cycle phosphorylation

7.2.3 The Krebs/Citric ATP ATP ATP

Acetyl CoA
Acid Cycle
NADH
+ H+ H2 O

NAD+
Oxaloacetate

Malate Citrate
Isocitrate

CO2
Citric
acid
cycle NAD+
H2 O
NADH
Fumarate + H+

α -Ketoglutarate

FADH2
CO2
NAD+
FAD
Succinate NADH
Pi
GTP GDP Succinyl + H+
CoA

ADP

ATP
Acetyl group of acetyl CoA
combines with OAA, forming
citrate.
Citrate regenerated back to OAA.
3 CO2 molecules released,
including one released during
conversion of pyruvate to acetyl
CoA.
Cycle generates one ATP per turn by
substrate-level phosphorylation.
GTP is formed by substrate-level
phosphorylation.
GTP is used to synthesize an ATP
directly.
Most of chemical energy
transferred to NAD+ and FAD
during redox reactions.
Reduced coenzymes NADH and
FADH2 then transfer high-energy
electrons to ETC.
For every acetyl CoA, each cycle
produces:

i. 1 ATP by substrate-level
phosphorylation
ii. 3 NADH, and
iii. 1 FADH2
Pyruvate
(from glycolysis,
2 molecules per glucose) Citric
CO2 Glycolysis
acid
Oxidation
phosphorylation
NAD+ cycle

CoA
NADH
ATP ATP ATP
+ H+ Acetyl CoA
CoA

CoA

Citric
acid 2 CO2
cycle
FADH2 3 NAD+

FAD 3 NADH
+ 3 H+
ADP + P i

ATP
Summary:
Acetyl CoA + 3 NAD+ + FAD + ADP + Pi
+2H2O

2 CO2 + CoA-SH + 3NADH + 3H+ +


FADH2 + ATP
Link
reactions
8.2.4 Electron transport chain
(oxidative phosphorylation)

4 out of 38 ATP are produced by


substrate-level phosphorylation:
Glycolysis – 2 ATP.

Citric acid cycle – 2 ATP.


NADH and FADH2 account for the
majority of energy extracted from food.
NADH and FADH2 link glycolysis and
citric acid cycle to oxidative
phosphorylation, which uses energy
released by ETC to power ATP
synthesis.
Pathway of Electron Transport

ETC is a collection of molecules


embedded in cristae.
Most components of ETC are
proteins bound to prosthetic
groups.
Electrons drop in free energy as they
pass down ETC.
NADH
50

FADH2

Multiprotein
I FAD complexes
40 FMN
Free energy (G) relative to O2 (kcal/mol)

Fe•S Fe•S II
Q
III
Cyt b Oxidative
Glycolysis Citric phosphorylation:
Fe•S acid electron transport
30 cycle
and chemiosmosis
Cyt c1 IV
Cyt c
Cyt a ATP ATP ATP

Cyt a3
20

10

0 2 H+ + 1/2 O2

H2O
During electron transport along ETC,
electron carriers alternate between reduced
and oxidized states as they accept and
donate electrons.

NADH
FAD
(reduced)
(oxidized)
CoQH2
Cyto. Oxidase
(reduced)
NAD (oxidized) H2O
FADH2
(oxidized)
(reduced)
CoQ Cyto. Oxidase H2
(oxidized) (reduced) ½ O2
Each component of chain becomes
reduced when it accepts electrons
from its “uphill” neighbor, which is
less electronegative.
It then returns to its oxidized form as
it passes electrons to its more
electronegative “downhill” neighbor.
Electrons carried by NADH are
transferred to the first molecule in
ETC, a flavoprotein.
Electrons continue along chain that
includes several cytochrome
proteins and one lipid carrier.
Prosthetic group of each
cytochrome is a heme group with
an iron atom that accepts and
donates electrons.
Last cytochrome of chain, cyt a3,
passes its electrons to oxygen, which
is very electronegative.
Each oxygen atom also picks up a
pair of H+ from aqueous solution to
form water.
For every two electron carriers (four
electrons), one O2 molecule is
reduced to two molecules of water.
Electrons carried by FADH2 have
lower free energy and are added
at a lower energy level than those
carried by NADH.
–ETC provides about one-third
less energy for ATP synthesis
when electron donor is FADH2
rather than NADH.
NADH
FADH2
ETC generates no ATP directly.
Its function is to break the large
free energy drop from food to
oxygen into a series of smaller
steps that release energy in
manageable amounts.
Chemiosmosis: Energy-Coupling
Mechanism
OXIDATIVE PHOSPHORYLATION: ETC &
CHEMIOSMOSIS : THE MECHANISM……
1. NADH delivers two electrons and two
protons to the first protein complex (I)
located in cristae.
2. Energy is released as the electrons
pass down the ETC (electrons move
from high energy to low energy level).
3. Energy is used to actively pump H+
across the membrane..
One of Its
A Mitochondrion Mitochondri
a a
A b
Cell
A
Crista
Outer
& Inner
Membrane
s c
ntermembran
e Matrix
4. Protons move from matrix to
intermembrane space
5. Protons are moved across by three (of
the four) complexes/complex I, III and IV
6. Proton gradient provide energy for ATP
synthesis.
7. Diffusion of protons from intermembrane
space to the matrix of mitochondrion
through ATP synthase (H+ only
permeable to ATP synthase)
8. As protons pass through it, energy is
obtained to phosphorylate ADP into ATP
ADP + Pi ATP.
9. After passing through three protein complexes,
electrons combine with one oxygen atom and two H+ to
form water (from matrix of mitochondrion)

2H+ + 1/2O2 + 2e H2 O

* Oxygen- final electron acceptor.


10. FADH2 delivers its electron via protein
complex II (at lower energy level).

1 FADH2 PRODUCES 2 ATP


1 NADH PRODUCES 3 ATP
3. Process is done two more time
times. Electrons are passed through
protein complexes III & IV, which
transport two more H+ across
membrane.
4. After passing through three protein
complexes, electrons combine with
one oxygen atom and two H+ to form
water.
Mitochondrial
(1)
5. This transport across membrane
produces a concentration gradient.
The H+ gradients that results is
referred to as a proton-motive
force. Gradient is used to make
ATP.
6. Cristae membrane impermeable to
H+ except through ATP synthase. As
protons pass through it, energy is
obtained to make ATP from ADP &
Pi.
7. FADH2 delivers its electron via
protein complex II – so, fewer
electrons are passed into inter-
membrane space.
8.2.5 Calculations of Total ATP Production
by Cellular Respiration
CYTOSOL Electron shuttles MITOCHONDRION
span membrane 2 NADH
or
2 FADH2

2 NADH 2 NADH 6 NADH 2 FADH2

Glycolysis Oxidative
2 2 Citric phosphorylation:
Glucose Pyruvate Acetyl acid electron transport
CoA cycle and
chemiosmosis

+ 2 ATP + 2 ATP + about 32 or 34 ATP


by substrate-level by substrate-level by oxidation phosphorylation, depending
phosphorylation phosphorylation on which shuttle transports electrons
form NADH in cytosol

About
Maximum per glucose: 36 or 38 ATP
Summary of
Glucose Metabolism
(2)
Products generated when a molecule of
glucose is oxidized to 6 CO2 molecules:

Conversions
NADH in cytoplasm produces 2 or 3 ATP
by oxidative phosphorylation depending
on shuttle system used to transport
electrons from cytosol into mitochondrion:
If electrons are passed to FAD, e.g.
brain cells - 2 ATP.
If electrons are passed to NAD+,
e.g., liver cells & heart cells = 3ATP
In mitochondria:
NADH - 3 ATP

FADH – 2ATP
2
FAD
NADH GP ETC
(glycolysis) FADH2

Malate-
NAD
NADH aspartate
ETC
(glycolysis) NADH
Glycolysis
Substrate-level phosphorylation = 2
ATP
2 NADH = 2 x 2 ATP = 4 ATP; or
= 2 x 3 ATP = 6 ATP

Formation of Acetyl CoA


2 NADH = 2 x 3 ATP = 6ATP
Krebs Cycle
6 NADH = 6 x 3 ATP = 18 ATP
2 FADH2 = 2 x 2 ATP = 4 ATP
Substrate-level phosphorylation = 2
ATP
Total Yield
Glycolysis = 2 ATP
Aerobic respiration = 34 or 36 ATP
5. Summary
Pathway Substrate-level Oxidative Total
phosphorylatio phosphorylation ATP
n

Glycolysis 2 ATP 2 NADH = 4 - 6 6-8


ATP
Acetyl CoA 2 NADH = 6 ATP 6
Krebs cycle 2 ATP 6 NADH = 18 ATP 24
2 FADH2 = 4 ATP

Total 4 ATP 32 – 34 ATP 36-38


Energy Harvested from Glucose
Cytoplasm) Glucose
2 Glycolysi 4
ATP s ATP
2 Pyruvates
Mitochondrial 2 NADH 2 CO2
Matrix) 2 NADH
6 NADH Krebs 4 CO2
2 FADH2 Cycle 2
ATP

(Inner
Membrane) Electron Water
Transport 32
Oxygen System ATP
Efficiency of respiration

Complete oxidation of glucose = 686


kcal/mol.
Phosphorylation of ADP to form ATP = 7.3
kcal/mol.
Efficiency of respiration
= 7.3 kcal/mol x 38 ATP/glucose x 100%
686 kcal/mol glucose
= 40%.
≈ 60% energy from glucose lost
as heat.
Some used to maintain body
temperature (37°C).
Efficient in energy conversion.
8.3 Anaerobic respiration
Qxidative phosphorylation ceases
in absence of O2.
Some cells oxidize organic fuel
and generate ATP without use of
O2 through fermentation.
E.g.,anaerobic catabolism of
sugars.
Fermentation generate ATP from
glucose by substrate-level
phosphorylation as long as there is
NAD+ to accept electrons.
If NAD+ pool is exhausted,
glycolysis shuts down.
Under aerobic conditions, NADH
transfers electrons to ETC, and
recycles NAD+.

Anaerobic - ATP generated by


glycolysis; NAD+ recycled by
transferring electrons from NADH
to pyruvate/derivatives of pyruvate.
8.3.1 Ethanol Fermentation

2 ADP + 2 P i 2 ATP

Glucose Glycolysis

2 Pyruvate

2 NAD+ 2 NADH 2 CO2


+ 2 H+

2 Ethanol 2 Acetaldehyde

Alcohol (ethanol) fermentation


Two steps:
1. Pyruvate converted to
acetaldehyde (2C), by removal of
CO2.
2. Acetaldehyde reduced by NADH
to ethanol.
Alcohol fermentation by yeast -
brewing and winemaking.
8.3.2 Lactic Fermentation

2 ADP + 2 P i 2 ATP

Glucose Glycolysis

2 NAD+ 2 NADH
+ 2 H+
2 Pyruvate

2 Lactate
Lactic acid fermentation
Pyruvate reduced by NADH to lactate
without release of CO2.
Lactic acid fermentation by fungi and
bacteria - cheese and yogurt.
Human muscle cells switch from aerobic
respiration to lactic acid fermentation to
generate ATP when O2 is scarce.
Waste product, lactate, causes muscle
fatigue - converted back to pyruvate in
liver.
GLUCOSE (6C)

2LACTATE (3C) 2ETHANOL (2C)


2NADH
+ 2H+
2NAD+

2NAD+

2NADH 2ETHANAL /
+ 2H+ ASETALDEHYDE (2C)
muscle cell/bacteria
2ATP CO2
2Pyruvates converted into
2lactate (3C) Yeast cell
2NADH are used 2Pyruvates converted into
2 ATP are produced 2ethanol (2C) &2CO2
2NADH are used
2PYRUVATES (3C) 2 ATP are produced
Fermentation and Cellular Respiration
Compared

Similarities – both use

1. Glycolysis : oxidize sugars to pyruvate


- 2 ATP produced by substrate-level
phosphorylation.
2. NAD+ : oxidizing agent - accept
electrons from food during glycolysis.
Difference
1. Mechanism for oxidizing NADH to NAD+.
 Fermentation - electrons of NADH
passed to an organic molecule to
regenerate NAD+.
 Respiration - electrons of NADH passed
to O2, generating ATP by oxidative
phosphorylation.
2. ATP generated per molecule of glucose.
 Aerobic : 36 - 38 ATP.

 Anaerobic : 2 ATP.
Facultative Anaerobes

Makes ATP aerobically if O2 is


present; switch to fermentation in
absence of O2.
E.g., yeast, many bacteria & human
muscle cells (cellular level).
Glucose

CYTOSOL

Pyruvate

No O2 present O2 present
Fermentation Cellular respiration

MITOCHONDRION

Ethanol Acetyl CoA


or
lactate
Citric
acid
cycle
8.4 Metabolism of Fat and Protein
Proteins Carbohydrates Fats

Amino Sugars Glycerol Fatty


acids acids

Glycolysis
Glucose

Glyceraldehyde-3-P

NH3 Pyruvate

Acetyl CoA

Citric
acid
cycle

Oxidative
phosphorylation
Catabolism

Carbohydrates:
Polysaccharides
(starch/glycogen) hydrolyzed to
glucose monomers that enter
glycolysis.
Hexose sugars (galactose and
fructose) - modified to undergo
glycolysis.
Protein:
Proteins digested to individual amino
acids.
– Amino groups removed via
deamination.
– Nitrogenous waste excreted as
ammonia, urea, or another waste
product.
Carbon skeletons modified by enzymes
and enter as intermediaries into
glycolysis or citric acid cycle.
Fats:
Fats digested to glycerol and fatty
acids.
Glycerol converted to G3P –
enters glycolysis.
Fatty acids split into 2C fragments
via beta oxidation.
Enter citric acid cycle as acetyl
CoA.
Biosynthesis (Anabolic Pathways)

Intermediaries in glycolysis and citric acid


cycle can be diverted to anabolic
pathways.
E.g., human cell synthesizes 10 different
amino acids by modifying compounds
from citric acid cycle.
Glucose synthesized from pyruvate.

Fatty acids from acetyl CoA.


Glycolysis and citric acid cycle
function as metabolic interchanges
that enable cells to convert one kind
of molecule to another.
E.g., conversion of excess proteins
and carbohydrates to fats through
intermediaries of glycolysis and citric
acid cycle.