Index to this page

 Mitochondria
Cellular Respiration  The Citric Acid Cycle
Cellular respiration is the process of oxidizing food  The Electron Transport Chain
molecules, like glucose, to carbon dioxide and water.
 Chemiosmosis in Mitochondria
C6H12O6 + 6O2 + 6H2O → 12H2O + 6 CO2
 How many ATPs?
The energy released is trapped in the form of ATP for use
by all the energy-consuming activities of the cell.  Mitochondrial DNA (mtDNA)

The process occurs in two phases:

 glycolysis, the breakdown of glucose to pyruvic acid

 the complete oxidation of pyruvic acid to carbon dioxide and water

In eukaryotes, glycolysis occurs in the cytosol. The remaining processes take place
in mitochondria.

Mitochondria

Mitochondria are membrane-enclosed organelles distributed through the cytosol of most

eukaryotic cells. Their number within the cell ranges from a few hundred to, in very active cells,
thousands. Their main function is the conversion of the potential energy of food molecules into
ATP.

Mitochondria have:

 an outer membrane that encloses the entire structure

 an inner membrane that encloses a fluid-filled matrix

 between the two is the intermembrane space

 The inner membrane is elaborately folded with shell-

like cristae projecting into the matrix.

 a small number (some 5–10) circular molecules of DNA

This electron micrograph (courtesy of Keith R. Porter) shows a single mitochondrion from a
bat pancreas cell. Note the double
membrane and the way the inner
membrane is folded into cristae. The dark,
membrane-bounded objects above the
mitochondrion are lysosomes.

The number of mitochondria in a cell can

  increase by their fission (e.g. following mitosis);

 Decrease by their fusing together.

(Defects in either process can produce serious, even fatal, illness.)

The Outer Membrane

The outer membrane contains many complexes of integral membrane proteins that form channels
through which a variety of molecules and ions move in and out of the mitochondrion.

The Inner Membrane

The inner membrane contains 5 complexes of integral membrane proteins:

 NADH dehydrogenase (Complex I)

 succinate dehydrogenase (Complex II)

 cytochrome c reductase (Complex III; also known as the cytochrome b-c 1 complex)

 cytochrome c oxidase (Complex IV)

 ATP synthase (Complex V)

The Matrix

The matrix contains a complex mixture of

soluble enzymes that catalyze the
respiration of pyruvic acid and other small
organic molecules.

Here pyruvic acid is

 oxidized by NAD+ producing NADH +

H+

 decarboxylated producing a
molecule of

o carbon dioxide (CO2) and

o a 2-carbon fragment of
acetate bound tocoenzyme
A forming acetyl-CoA

The Citric Acid Cycle

 This 2-carbon fragment is donated to a molecule ofoxaloacetic acid.

 The resulting molecule of citric acid (which gives its name to the process) undergoes the
series of enzymatic steps shown in the diagram.
  The final step regenerates a molecule of oxaloacetic acid and the cycle is ready to turn
again.

Summary:

 Each of the 3 carbon atoms present in the pyruvate that entered the mitochondrion leaves
as a molecule of carbon dioxide (CO2).

 At 4 steps, a pair of electrons (2e-) is removed and transferred to NAD+ reducing it

to NADH + H+.

 At one step, a pair of electrons is removed from succinic acid and reduces the prosthetic
group flavin adenine dinucleotide (FAD) to FADH2.

The electrons of NADH and FADH2 are transferred to the electron transport chain.

The Electron Transport Chain

The electron transport chain consists of 3 complexes of

integral membrane proteins

 the NADH dehydrogenase complex (I)

 the cytochrome c reductase complex (III)

 the cytochrome c oxidase complex (IV)

and two freely-diffusible molecules

 ubiquinone

 cytochrome c

that shuttle electrons from one complex to the next.

The electron transport chain accomplishes:

 the stepwise transfer of electrons

from NADH (and FADH2) to oxygen molecules to
form (with the aid of protons) water molecules
(H2O);

(Cytochrome c can only transfer one electron at a time, so

cytochrome c oxidase must wait until it has accumulated 4
of them before it can react with oxygen.)

 harnessing the energy released by this transfer to the pumping of protons (H+) from
the matrix to theintermembrane space.

 Approximately 20 protons are pumped into the intermembrane space as the 4 electrons
needed to reduce oxygen to water pass through the respiratory chain.
  The gradient of protons formed across the inner membrane by this process of active
transport forms a miniature battery.

 The protons can flow back down this gradient only by reentering the matrix through ATP
synthase, another complex (complex V) of 16 integral membrane proteins in the inner
membrane. The process is called chemiosmosis.

Chemiosmosis in mitochondria

The energy released as electrons pass down the gradient from NADH to oxygen is harnessed by
three enzyme complexes of the respiratory chain (I, III, and IV) to pump protons (H+) against their
concentration gradient from the matrix of the mitochondrion into the intermembrane space (an
example of active transport).

As their concentration increases there (which is the same as saying that the pH decreases), a
strong diffusion gradient is set up. The only exit for these protons is through the ATP
synthase complex. As inchloroplasts, the energy released as these protons flow down their
gradient is harnessed to the synthesis ofATP. The process is called chemiosmosis and is an
example of facilitated diffusion.

One-half of the 1997 Nobel Prize in Chemistry was awarded to Paul D. Boyer and John E. Walker
for their discovery of how ATP synthase works. Link to some of the details.

How many ATPs?

It is tempting to try to view the synthesis of ATP as a simple matter of stoichiometry (the fixed
ratios of reactants to products in a chemical reaction). But (with 3 exceptions) it is not.

Most of the ATP is generated by the proton gradient that develops across the inner mitochondrial
membrane. The number of protons pumped out as electrons drop from NADH through the
respiratory chain to oxygen is theoretically large enough to generate, as they return through ATP
synthase, 3 ATPs per electron pair (but only 2 ATPs for each pair donated by FADH 2).

With 12 pairs of electrons removed from each glucose molecule,

 10 by NAD+ (so 10x3=30); and

 2 by FADH2 (so 2x2=4),
this could generate 34 ATPs.

Add to this the 4 ATPs that are generated by the 3 exceptions and one arrives at 38.

But

 The energy stored in the proton gradient is also used for the active transport of several
molecules and ions through the inner mitochondrial membrane into the matrix.

 NADH is also used as reducing agent for many cellular reactions.

So the actual yield of ATP as mitochondria respire varies with conditions. It probably seldom
exceeds 30.

The three exceptions

A stoichiometric production of ATP does occur at:

 one step in the citric acid cycle yielding 2 ATPs for each glucose molecule. This step is the
conversion of alpha-ketoglutaric acid to succinic acid.

 at two steps in glycolysis yielding 2 ATPs for each glucose molecule.

Mitochondrial DNA (mtDNA)

The human mitochondrion contains 5–10 identical,

circular molecules of DNA. Each consists of 16,569
base pairs carrying the information for 37
genes which encode:

 2 different molecules of ribosomal

RNA (rRNA)

 22 different molecules of transfer

RNA (tRNA) (at least one for each amino
acid)

 13 polypeptides

The rRNA and tRNA molecules are used in the

machinery that synthesizes the 13 polypeptides.

The 13 polypeptides participate in building several

protein complexes embedded in the inner
mitochondrial membrane.

 7 subunits that make up the mitochondrial NADH dehydrogenase (complex I)

 cytochrome b, a subunit of cytochrome c reductase (complex III)

 3 subunits of cytochrome c oxidase (complex IV)

 2 subunits of ATP synthase (complex V)

Each of these protein complexes also requires subunits that are encoded by nuclear genes,
synthesized in the cytosol, and imported from the cytosol into the mitochondrion. Nuclear genes
also encode ~1,000 other proteins that must be imported into the mitochondrion. [More]

Mutations in mtDNA cause human diseases.

Mutations in 12 of the 13 polypeptide-encoding mitochondrial genes have been found to cause

human disease.

Although many different organs may be affected, disorders of the muscles and brain are the most
common. Perhaps this reflects the great demand for energy of both these organs. (Although
representing only ~2% of our body weight, the brain consumes ~20% of the energy produced
when we are at rest.)

Some of these disorders are inherited in the germline. In every case, the mutant gene is received
from the mother because none of the mitochondria in sperm survives in the fertilized egg. Other
disorders are somatic; that is, the mutation occurs in the somatic tissues of the individual.

Example: exercise intolerance

A number of humans who suffer from easily-fatigued muscles turn out to have a mutations in
theircytochrome b gene. Curiously, only the mitochondria in their muscles have the mutation; the
mtDNA of their other tissues is normal. Presumably, very early in their embryonic development, a
mutation occurred in a cytochrome b gene in the mitochondrion of a cell destined to produce their
muscles.

The severity of mitochondrial diseases varies greatly. The reason for this is probably the extensive
mixing of mutant DNA and normal DNA in the mitochondria as they fuse with one another. A
mixture of both is called heteroplasmy. The higher the ratio of mutant to normal, the greater the
severity of the disease. In fact by chance alone, cells can on occasion end up with all their
mitochondria carrying all-mutant genomes — a condition called homoplasmy (a phenomenon
resembling genetic drift).

Mutations in some 228 nuclear genes have also been implicated in human mitochondrial diseases.

Why do mitochondria have their own genome?

Many of the features of the mitochondrial genetic system resemble those found in bacteria. This
has strengthened the theory that mitochondria are the evolutionary descendants of a bacterium
that established an endosymbiotic relationship with the ancestors of eukaryotic cells early in the
history of life on earth. However, many of the genes needed for mitochondrial function have since
moved to the nuclear genome.

The recent sequencing of the complete genome of  Rickettsia prowazekii has revealed a number
of genes closely related to those found in mitochondria. Perhaps rickettsias are the closest living
descendants of the endosymbionts that became the mitochondria of eukaryotes.