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Lec-Aeo-11-Biochem 2018
Lec-Aeo-11-Biochem 2018
Substrates possessing higher redox potential enter the electron transfer chain through flavoprotein
dehydrogenase - complex II. It transports the electrons to coenzyme Q. Complex II contains FAD and Fe-S
(iron-sulfur) complexes. Complex III is Co-Q - cytochrome C reductase. It consist of cytochromes b and c1,
and 2Fe-2S iron sulfur clusters.
The cytochrom C is a soluble small (12 kD) hemoprotein. It carries electrons to complex IV - cytochrome
oxidase.
Cytochrome oxidase contains two cytochromes: a and a3 and two copper ions. The cytochrome oxidase
reaction accounts for more than 90% of all oxygen consumed by humans. The reduction of oxygen by
cytochrome oxidase accounts for production of metabolic water of about 300 ml per day.
The enzymes or complexes of electron transfer chain are distributed in anisothropic way: the first, third and
fourth complexes are incorporated transmembraneously. The second complex is distributed on the inner
side.
The connection between the first and the third complexes is performed by Co-Q, lipophylic molecule which
moves easily inside the inner membrane.
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The hydrogen transport between complexes II and III is mediated in the same way, by CoQ. The soluble
cyt.C transports electrons from third complex to the fourth. It is the only soluble and non-fixed cytochrome
in electron transfer chain.
The transfer of hydrogen atoms and electrons
proceeds with energy release. These energy is
released in portions and sustains a synthesis of
ATP, which occurs in complex V - ATP-synthase.
Both processes are coupled: oxidation insures the
energy for ATP synthesis.
2. ATP-synthase structure. ATP - synthase is an
aggregate of polypeptides that form a mushroom-
shaped molecule on the inner side of the inner
mitochondrial membrane.
The enzyme abbreviation is FoF1 - ATP -ase. Fo is
oligomycin sensitive subunit. It is embedded in
the membrane and functions as a proton pump. F1
subunit is composed of several polypeptide chains,
designated αβδεγ. It forms a mushroom top.
When separated, F1 possesses ATP hydrolysing activity, i.e. ATP-ase activity. The composition of mammalian
mitochondrial enzyme is α 3 β3γδε1-2. The triplet of α3β3 subunits provides three ATP binding sites per
molecule.
3. Inhibition of electron transfer chain.
The first complex is inhibited by rotenone, amytal and barbiturates. Rotenone is an exotic poison which was
used by Indians in fishing, and barbiturates are in sleeping pill-soporific.
Inhibitors accept electrons and stop their flow through electron transfer chain. The consumption of oxygen
stops. If the electron transfer chain is procured with
substrates for complex II, its function will be
restored.
Complex II is inhibited by carboxin and TTFA [4,4,4-
trifluoro-1-(2-thienyl)-1,3-butanedione].
The third complex is inhibited by the antibiotic
antimycin. The block in vitro may be recovered by
vit.C which serves as electron donor for complex IV -
cytochrome oxidase. Cytochrome oxidase is inhibited
by (CN-) cyanides, carbonmonooxide and by azides.
All these compounds bind (Fe) ferrous ions in the
cytochrome and prevent electron transfer.
The ATP - synthase is inhibited by the antibiotic
oligomycin. It completely inhibits oxidative
phosphorylation. Atractyloside is another antibiotic
which inhibits ADP/ATP transport through
mitochondrial membrane.
The ATP production. When electron transfer chain
works from the first complex i.e. NADH -DH, it produces 3 ATP molecules on transferring of 2 hydrogen
atoms or electrons through electron transfer chain.
In case it uses the other “entrance” - complex II,
and starts from FADH2, the electron transfer chain
produces 2ATP molecules only. When electrons
enter the complex IV, the electron transfer chain
produces only one ATP molecule (physiological
substrates for complex IV are not known).
The difference in ATP production may be explained
by the difference in total redox potential between
substrate and oxygen pairs.
For NADH and O2 pair the redox potential equals 1.24
V (Ered. = + 1.24 V).
For FADH2 and O2 pair the redox potential equals
1.06 (Ered. = +1.06).
For cyt.oxidase and O2 pair the redox potential -
Ered. = 0.60 v.
In fact, the number of ATP molecules formed from ADP and Pi per electron pair transported to an atom of
oxygen ( 1/2 O2 ) is termed P /O ratio. With NADH as electron donor the P/O ratio is between 3 and 2.
Current data suggest that it is 2.5.
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With succinate as electron donor, P/O ratio is 1.5, that is from a mole of oxidated succinate 1.5 moles ATP
are produced.
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complex IV and cyt c the electron transfer chain is active, if its relation lower, the electron transfer chain
is regulated by feedback control. At high ATP levels, ADP is insufficient for ATP-ase, the phosphorylation
stops. High levels of transmembrane potential impede/block up the oxidation of NADH in electron transfer
chain.
Substrate oxidation in the TCA cycle activates the respiratory chain, leading to the generation of ATP, which
is subsequently translocated to the cytosol.
ANT, adenine nucleotide translocator; GDH, glutamate dehydrogenase; Glu, glutamate; KG, ketoglutarate;
OAA, oxaloacetate; PC, pyruvate carboxylase; PDH, pyruvate dehydrogenase; Suc-CoA, succinyl-CoA; SDH,
succinate dehydrogenase; TCA, tricarboxylic acid; UCP2, uncoupling protein 2.
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