MITOCHONDRIA

CELL BIOLOGY

1
 Endosymbiosis Hypothesis
A A prokaryote ingested some aerobic bacteria. The
aerobes were protected and produced energy for the
prokaryote

A B C D

Cyanobacteria
Aerobic bacteria Mitochondria Chloroplasts

N
N N

Prokaryote Plant cell

N
Animal Cell
 Endosymbiosis Hypothesis

B Over a long period of time the aerobes became

mitochondria, no longer able to live on their own

A B C D

Cyanobacteria
Aerobic bacteria Mitochondria Chloroplasts

N
N N

Prokaryote Plant cell

N
Animal Cell
 Endosymbiosis Hypothesis

C Some primitive prokaryotes also ingested

cyanobacteria, which contain photosynthetic
pigments

A B C D

Cyanobacteria
Aerobic bacteria Mitochondria Chloroplasts

N
N N

Prokaryote Plant cell

N
Animal Cell
 Endosymbiosis Hypothesis

D Cyanobacteria became chloroplasts, unable to live

on their own

A B C D

Cyanobacteria
Aerobic bacteria Mitochondria Chloroplasts

N
N N

Prokaryote Plant cell

N
Animal Cell
 Endosymbiosis

Mitochondria formed as a
result of an endosymbiotic
event around 2 billion years
ago.
Mitochondria: Structure and
Function
 First identified in animals in 1840, then in plants in
1900
 Have an inner and outer phospholipid membrane making
up the envelope
 Outer membrane smooth, inner membrane folded into
cristae for a large surface area
 Space between the inner and outer membrane known as
the intermembrane space
 The matrix is the middle bit (inside the inner
membrane) it is gel like and made of proteins and
lipids, looped mitochondrial DNA ribosomes and
enzymes
Structure and Function
 The matrix is where the link reaction and
Krebs cycle take place- it contains:
 Enzymes that catalyse these stages
 NAD molecules
 Oxaloacetate
 Mitochondrial DNA that codes for
mitochondrial proteins
 Mitochondrial ribosomes (like bacterial
ribosomes)
Relative contributions of nuclear
and mitochondrial genes to protein
composition
Mitochondria are
organized into 4 distinct
compartments
 Outer membrane:
 Perforated with large
channels (porins) that
allow entry of molecules <
5000 kD
 Phospholids with proteins
forming channels allowing
pyruvate through
 Enzymes involved in
mitochondrial lipid
synthesis
 Mitochondria are
organized into 4 distinct

compartments
Intermembrane space:

 Compartment into which H+ is

pumped
 Enzymes that use newly-made ATP
to phosphorylate other nucleotides
 Mitochondria are
organized into 4 distinct
Innercompartments
membrane:
 Different lipid composition from outer
membrane
 Impermeable to most small ions including
Hydrogen ions (protons)
 Folded into cristae to maximize surface area
 Proteins that carry out redox reactions of the
electron transport chain
 Proteins that synthesize ATP
 Transport proteins that move molecules into
and out of the matrix
 Electron carriers and ATP synthase embedded
into it
 Mitochondria are
organized into 4 distinct
 compartments
Matrix:

 Internal space containing

enzymes for Krebs cycle
 Contains mitochondrial
DNA, special ribosomes,
tRNAs, and enzymes
required for gene
expression
Mitochondria use pyruvate
or fatty acids to make energy

Pyruvate from sugars, fatty acids from fats

High energy electrons are
generated via the citric acid
(Krebs) cycle
Mitochondria & ATP
 Thetheoretical maximum yield of
ATP per molecule of glucose is
rarely, if ever, achieved in aerobic
respiration;
 howthe structure of mitochondria
enables them to carry out their
functions;
Yield of ATP
 We have seen that during the electron
transport chain, most ATP is made (by
substrate level phosphorylation)
 Together with the ATP made during glycolysis
and the Krebs cycle, the total yield of ATP
molecules, per molecule of glucose respired,
should be 30
 However, this is only a theoretical yield, in
real situations the maximum yield (amount
made) of ATP is not always possible
 why the maximum yield of ATP is rarely
achieved
 According to some of newer sources the ATP yield
during aerobic respiration is not 36–38, but only
about30–32
 ATP molecules / 1 molecule of glucose, because: ATP :
NADH+H+ and ATP : FADH2 ratios during the oxidative
phosphorylation appear to be not 3 and 2, but 2.5 and
1.5 respectively

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 Complete Glucose Oxidation follows :
 Glycolysis (Glucose > 2Pyruvic Acid) - Here 2 ATPs are produced by substrate level phosphorylation and 2 NADH are also produced .
 Transition Step ( Pyruvic acid> Acetyl CoA). - Here 1 NADH is produced in this step. (But we had 2 Pyruvic Acid . So we have two
2NADH and 2Acetyl CoA produced in these steps)
 Krebs Cycle (This cycle will utilise 1 Acetyl CoA and will produce 1GTP, 3NADH & 1FADH2). But here again we have to oxidise
complete 2 Acetyl CoA. So , 2GTP (equivalent to 2 ATP), 6NADH, 2FADH2.
 Calculation:
 ATP : 2+2 = 4
 NADH : 2 +2+6 = 10
 FADH2 : 24 . Last step in Glucose Oxidation is Oxidative Phosphorylation.
 Here ,
 1NADH after passing through electron carriers result in 3 ATP .
 1FADH2 after passing through electron carriers result in 2 ATP.
 So, 10 NADH = 30 ATP & 2 FADH2 = 4 ATP.
 Again totaling ,
 ATP : 2+2 = 4
 NADH : 2 +2+6 = 10 ~ 30 ATP
 FADH2 : 2 ~ 4ATP
 NET : 38 ATP.
 So , when 1 Glucose undergoes aerobic respiration , net 38 ATP are produced.

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 Oxidation of glucose through the aerobic respiration gives
the ATP following...
 Glycolysis- 2 ATP, 2 NADPHH+

 Oxidative decarboxylation- 2 NADPHH+

 Citric acid cycle/ Kreb's cycle - 2 ATP, 6 NADPHH+, 2

FADH2

 Through Electron Transport Chain

 NADHH+ gives 2.5 ATP,
 FADH2 gives 1.5 ATP
 So totally we get the 32 ATP

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An overview of carbohydrate metabolism in eukaryotic cells
 Some protons leak
across the
mitochondrial
membrane, reducing
the number of
protons to generate
the proton motive
force

Some ATP is used for

the shuttle to bring Some ATP produced
Hydrogen from is used to actively
reduced NAD made transport pyruvate
during glycolysis, in into the
the cytoplasm, into mitochondria
the mitochondria
24
ATP Synthase

 Large and protrude from inner membrane

into matrix
 Known as stalked particles
 Allow protons through (H+)

ATP sythase is a protein complex

embedded in the inner mitochondrial
membrane
Phosphorylation

The F1Fo ATPase

(or ATP synthase)
is a molecular motor

-it uses the PMF to

make ATP

-it can also be

reversed (using ATP
hydrolysis to
recharge the PMF)
ATP synthase acts as a rotary
motor
 Proton motive force (PMF)
The PMF is an electrochemical gradient of membrane
potential (ΔΨ) and pH (ΔpH)
 Electron Carriers
 Enzymes with non protein haem cofactors
(containing iron)
 The iron atoms become reduced Fe3+ to
Fe2+ by accepting an electron (e-) then re-
oxidised to Fe3+ by passing the electron
onto the next carrier
 Oxidoreductase enzymes are involved in the
oxidation and reduction reactions
 Electron carriers also have a coenzyme that
pumps hydrogen ions from the matrix into
the intermembrane space
Mitochondria catalyze a major conversion
of energy by oxidative phosphorylation

Text
 Mitochondria make other products

Mitochondria produce biosynthetic precursors

OXPHOS also leads to the production of:

•Superoxide:formed when O2 steals electrons

from the ETC complexes

•Heat:
a by-product of the reactions of
OXPHOS
 Oxidation and Electron Transport
Electrons from NADH and FADH2 are passed down
respiratory chain to O2

Electron transport expels protons, creating a proton

gradient- the proton motive force (PMF)
Oxidation and phosphorylation are coupled
by a shared dependence on the PMF
 Uncoupling proteins
Many mammals warm vital tissues using brown fat

Adipose tissue with abundant mitochondria that

possess a the protein thermogenin (or uncoupling
protein 1).

UCP-1 short-circuits the proton gradient,

increasing VO2 and heat production.

All eukaryotes have proteins related to UCPs, that

are thought to prevent the PMF from “over-
charging”, thereby reducing ROS production.
Protons are pumped across the inner
mitochondrial membrane
The electron transport chain consists
of 3 enzyme complexes
The electrochemical gradient of H+
across the inner membrane has 2
components:
The proton gradient drives ATP
synthesis

Text
The proton gradient also
drives coupled transport
Mitochondria The organelle that releases
energy in the cell. (The powerhouse of the
cell)
Mitochondria produce ATP using energy
stored in food molecules.
 Mitochondria are the primary energy producers in cells.

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Structure

 Mitochondria have a double membrane structure

 There is a single outer membrane and a folded inner

membrane

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 Sac with two inner compartments which are separated
by the inner membrane.

 The first compartment is between the outer and inner

membranes.

 The outer compartment is inside the inner membrane.

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 The outer mitochondrial membrane is composed of
about 50% phospholipids by weight and contains a
variety of enzymes involved in such diverse activities as
the elongation of fatty acids, oxidation of epinephrine
(adrenaline), and the degradation of tryptophan.

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 The inner membrane contains proteins
with three types of functions [Alberts,
1994]:
 those that carry out the oxidation
reactions of the respiratory chain
 ATP synthase, which makes ATP in the
matrix
 specific transport proteins that
regulate the passage of metabolites
into and out of the matrix.
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Function

 Mitochondria are the site of most of the energy

production in eukaryotic cells .

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 They use complex molecules and oxygen to produce a
high energy molecule know as ATP (Adenosine
Triphosphate)
 process called aerobic respiration

47
 Energy production the mitochondria has been called the
"powerhouse of the cell".

48
 Mitochondria are very abundant in cells that require lots
of energy.
 Ex:- Muscle

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Unique

 Mitochondria are very unique in several regards

 have their own circular DNA
 have their own Ribosomes.

(The DNA in the cell nucleus does not code for the
construction of mitochondria. )

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 All the mitochondria in your body came from your
mother.
 Mitochondria are not part of the genetic code in the
nucleus of your cells.

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 Fathers only give genes to their children.

 Mothers give genes and cytoplasm to their children in their egg cells.

 Since mitochondria are in the cytoplasm and reproduce themselves they

only are inherited from mothers

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 Geneticists have used this curious feature of
mitochondria to study maternal family lines and rates of
evolution.

53
 Although the primary function of mitochondria is to convert organic materials into
cellular energy in the form of ATP, mitochondria play an important role in many
metabolic tasks, such as:
 Apoptosis-Programmed cell death
 Glutamate-mediated excitotoxic neuronal injury
 Cellular proliferation
 Regulation of the cellular redox state
 Heme synthesis
 Steroid synthesis

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 Heat production (enabling the
organism to stay warm).
 Some mitochondrial functions are
performed only in specific types of
cells. For example, mitochondria in
liver cells contain enzymes that allow
them to detoxify ammonia, a waste
product of protein metabolism. A
mutation in the genes regulating any
of these functions can result in a
variety of mitochondrial diseases.
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 Energy Conversion

 Mitochondria convert the chemical energy of reduced carbon

compounds into ATP.
 Chloroplasts convert light energy to the energy of reduced
carbon compounds
 Fundamental to both processes is an electron transport chain,
where energy is passed from compound to compound in
coupled oxidation:reduction reactions
2 membranes
Outer Membrane - has large Mitochondria
aqueous channels made of a
protein called porin -
permeable to most small
molecules
Inner Membrane - impermeable
to ions and small molecules
except by specific membrane-
transport proteins
 2 compartments
 MATRIX - enzymes -
TCA cycle, DNA -
Mitochondria several copies,
ribosomes, tRNA, etc
 INTERMEMBRANE
SPACE
Inner Membrane is folded
into cristae,that greatly
increase its surface
area. This is where you
find the Electron
Transport Chain (ETC),
ATP synthase, transport
proteins to allow
substances in to matrix
Fig 1-11
 Mitochondrial Structure
A How many compartments
do mitochondria have?
Which letter corresponds
to:
C • the matrix?
• a crista?
• the intermembrane
space?
E
• the outer membrane?
• the inner membrane?
Where would you find the
D DNA? the ribosomes?

B
 ELECTRON TRANSPORT CHAIN:

• The ETC is found on the inner membrane

• High energy electrons are passed along the ETC
generating a proton gradient across the inner
membrane
• H+ gradient drives ATP synthesis
1. Mitochondrial membranes

 The outer membrane is thought to be homologous to

an outer membrane present as part of the cell wall of
certain bacterial cells.
 The inner membrane is highly impermeable; all
molecules and ions require special membrane
transporters to gain entrance to the matrix.
 Possess ribosomes, circular DNA to manufacture their
own RNAs and proteins
The glycerol phosphate
shuttle
 Electrons are transferred from NADH to
dihydroxyacetone phosphate (DHAP) to form glycerol 3-
phosphate, which shuttles them into the mitochondrion.
These electrons then reduce FAD at the inner
membrane, forming FADH2 which can transfer the
electrons to a carrier of the electron-transport chain.
An overview of carbohydrate metabolism in eukaryotic cells
 2. The importance of reduced
coenzymes in the formation of ATP
(Chemiosmosis)
 1. High-energy electrons are passed
from FADH2 or NADH to the first of a
series of electron carriers in the
electron transport chain.
 2. The controlled movement of
protons back across the membrane
through an ATP-synthesizing enzyme
provides the energy required to form
ATP from ADP.
 A summary of the process of oxidative
phosphorylation
5.3 The role of mitochondria
in the formation of ATP
 1. Electron transport
 2. Oxidation-reduction potentials
 3. Types of electron carriers
1. Oxidation –reduction potential

Oxidizing agents can be ranked in a

the greater the affinity, the stronger the oxidizing agent.
series according to their affinity for
electrons:
 Reducing agents can also be ranked
according to their affinity for
electrons:
 The lower the affinity, the stronger the reducing agent
 Reducing agents are ranked according to electron-
transfer potential, such as NADH is strong reducing
agent, whereas those with low electron-transfer
potential such as H2O, are weak reducing agents.
Oxidizing and reducing agents occur as
couples such as NAD+ and NADH.

Strong reducing agents are coupled to

The affinity of substances for
electrons can be measured by
instruments that detect voltage—
oxidation-reduction (redox)
potential.
2. Electron transport

 1. Five of the nine reactions in matrix in Fig.

5.7 are catalyzed by dehydrogenases that
transfer pairs of electrons from substrates to
coenzymes, NADH and FADH2 → electron-transport
chain

 2. NADH and FADH2 dehydrogenase are

located in the inner membrane of
mitochondria.
3. Types of electron carriers

 Flavoproteins
 Cytochromes
 Ubiquinone
 Iron-sulfur proteins
Electron-transport complexes

 1. Complexes I, II, III, IV ----Fixed in

place
 2. I, III, VI in which the transfer of
electrons is accompanied by a
major release of free energy.
 2. Ubiquinone (lipid-soluble),
cytochrome c (soluble protein in the
intermembrane space)----move
within or along the membrane
I, III, VI are described as proton pump which drive the production of ATP.
5.5 The mechinery for ATP
formation
 1. The structure of ATP synthase
 2. The basis of ATP formation according to the binding
change mechanism
 3. Other roles for the proton-motive force in addition to
ATP synthesis