The Architecture of

Mitochondria is
Essential for
Mitochondrial Function

Gwen V. Childs, Ph.D.

Professor and Chair
Neurobiology and Developmental Sciences
501 686-7020; childsgwenv@uams.edu
http://cytochemistry.net/cell-biology/medical/

“…from the United Mitochondrial

Diseases Foundation”

“Imagine a major city with half its power plants

shut down. At best, such conditions would
produce a "brown out" with large sections of the
city working far below optimum efficiency.

Apply this to your body:

“Now imagine your body with one-half of its
energy producing facilities shut down. The brain
would be impaired, vision would be dim, muscles
would twitch spastically or would be too weak to
allow your body to walk or write, your heart would
be weakened, and you would not be able to eat
and digest your food.”

“For large numbers of people, especially

children, this is precisely the situation in which
they find themselves due to defects in the
mitochondria…. Mitochondrial diseases
compromise their lives and can be fatal.”

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 “….think mitochondria”
This is the watchword of the Mitochondrial
foundation. They recommend activating
this rule of thumb whenever:

z A "common disease" has atypical features that

set it apart from the pack.
z Three or more organ systems are involved.

z Recurrent setbacks or flare ups in a chronic

disease occur with infections.

Nervous system: Seizures, tremors, developmental delays,

dementia, stroke before age 40, poor balance, problems with
peripheral nerves.
Heart: cardiomyopathy, heart failure, conduction block.
Liver: liver failure, especially in babies
Kidneys: Faconi syndrome (loss of essential metabolites in urine).
Eyes: Drooping eyelids (ptosis); inability to move eyes from side
to side (external opthalmoplegia). Blindness (retinitis
pigmentosis).
Skeletal Muscle; Muscle weakness, exercise intolerance, cramps.
Digestive tract: acid reflux, vomiting, diarrhea, intestinal obstructi
Pancreas: Diabetes.
Widespread
Widespreadeffects
effectsof
ofmitochondrial
mitochondrialdiseases.
diseases. Most
Most
are
areinherited!
inherited! Mendelian
Mendelian oror Maternal!
Maternal!

Objectives of this
presentation:
Correlate structure with function
z Mitochondrial Geography
z Where are different enzyme systems
located
How do mitochondria reproduce
themselves?
z How do proteins enter?
z Mitochondrial inheritance
z Diseases involving mitochondria

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 Mitochondrial Compartments
Cristae mitochondriales Outer membrane

matrix

ribosomes
DNA Inner membrane

Mitochondrial structure
Identify the
regions in a
mitochondrion

What systems
are found in
each region?

How are mitochondria organized

to be powerhouses?
Sugars

Amino Pyruvate
Acids
Acetyl Kreb’s cycle
Fatty Co-A
acids

Glucose converted to pyruvate

by glycolysis. (Produces only
two molecules of ATP). CO2
Pyruvate carried into matrix
and converted to
Acetyl Co-A, forming CO2

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 How are mitochondria organized
to be powerhouses?
Sugars

Amino Pyruvate
Acids
Acetyl Kreb’s cycle
Fatty Co-A
acids
Acetyl Co-A joins with
oxaloacetic acid and
thus begins the Kreb’s Where does our
cycle CO2 CO originate?
2

Products of Kreb’s cycle? Electrons (H+ ions) + CO2

What happens to the

electrons?
Electrons are then carried from Kreb’s cycle to
Electron Transport Chain in cristae,
Two carrier molecules transport them to the
chain: nicotinamide adenine dinucleotide
(NAD+) and flavin adenine dinucleotide
(FAD+)
NAD+ + H+ NADH
FAD+ + 2 H+ FADH2
There, they are pumped with hydrogen pumps
to the space between the membranes.

Kreb’s Cycle/Citric Acid cycle

All but dehydrogenases at step 5 and 7 are in the matrix.

Alpha ketoglutarate dehydrogenase and succinate dehydrogenase are
Integral proteins in the inner membrane with active site facing the matrix.

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 Summary of how each step in the process produces
the carriers and C02
However, if there wasn’t a system to regenerate NAD,
the supply would dwindle.

Lodish et al, Molecular Cell biology, 5th ed.

Regeneration of NAD+
NAD+ in cytosol picks up electrons
during glycolysis and is converted to
NADH.
NADH cannot be transported across
mitochondrial inner membrane.
So, to get the electrons from the
cytoplasm across to the electron
transport chain, the mitochondria use
the malate shuttle.
In the process, NAD+ is restored in the
cytoplasm.

Aspartate gives up amino

group to alpha-ketoglutarate. Gives H+ to Oxaloacetate to form malate

Antiporter
transfers
malate
In exchange
Impermeable for alpha-
to oxaloacetate ketoglutarate
Antiporter transfers aspartate out in exchange for
glutamate

Oxaloacetate gets amino group

from glutamate to form aspartate. Malate gives up H+ to form oxaloacetate

Malate shuttle
Lodish et al, Molecular Cell biology, 5th ed.

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 Electron Transport chain on NADH and FADH2
Cristae membrane from Kreb’s cycle

Cristae
e-
Passes electron e- to
Electron transport chain
H+
Inner
Matrix As electron is passed
H+ down, the complexes pump
membrane H+ out of matrix
H+
ADP + Pi
ATP H+
synthase ATP

Outer Large electrochemical proton

Where is membrane gradient drives ATP synthase
ATP produced? to convert ADP to ATP

Electron transport chain

Intermembrane space: e- transport chain sets up
H+ gradient nH+
nH+ nH+
Cytochrome C

NADH
Dehydrogenase Cytochrome
complex Oxidase
Inner complex
membrane
e- B-C1 complex
ubiquinone
2H+ +
NADH ½ O2
NAD
+ H+
Matrix Negative charges

H2O

Cartoon showing
process

http://bcs.whfreeman.com/lodish5e/
z Log on; use narrated version to show
details.

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 Cytochrome C
histochemistry •Cytochrome C
lies near outer
leaflet of cristae
membrane.

•This is shown
by
histochemical
reaction.

4 electrons
released
from 4
molecules of
Cyt C

+ 4 protons
Join with
one O2
molecule to
form 2
molecules of
H2 O

Lodish et al, Molecular Cell biology, 5th ed.

Trace the packets of

energy
Glucose has energy in chemical bonds of about 680
kcal/molecule.
90% of this is conserved in the NADH and FADH2
Mitochondrial architecture allows by step by step transfer
of electrons along the transport chain which are close
together (packed) in the cristae.
Each transfer allows the energy to be released in small
packets and stored as the “proton-motive” force.
z Moving the electrons along drives the proton pumps which
sets up the concentration gradient as well as the electrical
potential
z These two ingredients are vital for the final transfer of the
energy packet to ATP

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 H+ H+ H+
ATP
H+ H+ synthase
nH+

F0 Inner
Membrane

nH+
Matrix

F1
Elementary
Particle
ADP + Pi ATP (F0, F1 particle)

Production of ATP

http://bcs.whfreeman.com/lodish5e/
z Log on; use narrated version to show
details.

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 Export of ATP and metabolites
from mitochondria
Same Proton motive force that powers the ATP
synthase also powers transporters for exchange of
pyruvate, malate, aspartate and glutamate.
Also powers the ATP/ADP antiporter coupled to the
phosphate antiporter.
z Phosphate transporter catalyzes import of phosphate
coupled to the export of OH
z OH binds proton to form water.
z ATP/ADP antiporter is highly abundant and exchanges one
ADP for one ATP
For every 5 protons pumped into the intercristal
space, 4 are used to make ATP and one is used to
power its export.

Elementary particles
ATP synthetase

Where is ATP produced??

Is it normal to see
lots of mitochondria
or cristae in the
mitochondria?

In some regions of the body, or

some types of tissue, yes.
z Hummingbird flight muscle [allow
wing beats of 80/sec (200/sec during
courtship)]
z Regions near sites of active
transport (high energy-ATP needs).

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 Hummingbird
Flight muscle
Cross-section
Note lots of
mitochondria!

Longitudinal section, skeletal

Muscle (mammal)

Epithelia: basolateral surface specialized for active transport .

Mitochondria respond to needs by replicating

In the region where ATP is needed

Mitochondrial replication
Like their bacterial precursors,
mitochondria replicate their DNA and
divide by splitting to form two daughter
mitochondria.
Mitochondria DNA replication and
division occurs during interphase,
before the nuclear DNA replication.
Mitochondria replicate when and where
needed. Feedback may be a drop in
ATP.

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 Mitochondrial
replication

What can mitochondrial DNA encode?

Human mtDNA is 16,569 bp

Encodes a number of mitochondrial
proteins
z Subunits 1, 2, and 3 of cytochrome
oxidase
z Subunits 6, 8,9 of the Fo ATPase
z Apocytochrome b subunit of CoQH2-
Cytochrome C reductase
z Seven NADH-CoQ reductase subunits

ATP Synthase

Poulton, J and Marchington DR, Reproduction 121: 751-755 (2002)

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 Mitochondrial DNA: circular, similar to bacteria

Mitochondrial DNA: There may be 10-12 copies

Of DNA/mitochondrion

What RNA’s are in mitochondria?

22 tRNAs
rRNA’s
z 16S
z 12S
z 5S1
___________
1Magalhaes, PJ; Andreu, AL, Schon EA, Evidence for the
presence of 5S rRNA in mammalian mitochondria Mol Biol
Cell 9: 2375-2382.

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Mitochondrial ribosomes

Mitochondrial organizing
center

Experiments with Yeast

mitochondria
Sorgo and Yaffe, J Cell Bio. 126: 1361-1373,
1994.
z showed the result of the removal of an outer
membrane protein from mitochondria called
MDM10 (from yeast).
z The mitochondria are able to take in
components and produce membranes and
matrix enzymes. However, fission is not
allowed.
z the result is a giant mitochondrion in each
yeast cell.

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 Yeast Giant
Mitochondria

Mutant lacking
mdm10-1 protein
contains one very
large
mitochondrion

When is replication not normal?

Mitochondria fail to make ATP, this
signals the cell to make more
Proliferation is often a sign of
mitochondrial disease…attempts to
compensate for inadequate supplies of
energy stores
Proliferation may be accompanied by
production of lots of
cristae…mitochondria may seem very
“dark”. Again, this is a compensatory
response.

•Unusual mitochondria in child with

mitochondrial disease
•Note many cristae on the right as if this
organelle is trying to compensate for its lack of
function.

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