4/29/2024

Complexes of electron
transport chain
Assignment: 02

Course: Bioenergetics
Department: FACULTY OF LIFE SCIENCES AND
INFORMATION TECHNOLOGY

Submitted To: Ma’am Noshaba mehmood

SUBMITTED BY: HUMAIRA-L1F20BSBC0030
Topic:

Complexes of electron transport chain, their compositions

and flow of electrons through the complexes and how ATP
formed in electron transport chain

Introduction to Electron Transport Chain (ETC):

The electron transport chain (ETC) is a series of protein complexes located in the inner
mitochondrial membrane (or the plasma membrane in prokaryotes). Its primary function is to
transfer electrons from electron donors (such as NADH or FADH2) to electron acceptors (such
as oxygen), generating a proton gradient across the membrane. This gradient is utilized by ATP
synthase to produce ATP via oxidative phosphorylation, a key process in cellular energy
production [1].

Complexes of the Electron Transport Chain:

 Complex I (NADH-CoQ Reductase):

Composition:

Complex I, also known as ubiquinone oxidoreductase, is made up of NADH dehydrogenase,

flavin mononucleotide (FMN), and eight iron-sulfur (Fe-S) clusters. The NADH donated from
glycolysis, and the citric acid cycle is oxidized here, transferring 2 electrons from NADH to
FMN. Then they are transferred to the Fe-S clusters and finally from Fe-S to coenzyme Q.
During this process, 4 hydrogen ions pass from the mitochondrial matrix to the intermembrane
space, contributing to the electrochemical gradient. Complex I may also play an important role
in causing apoptosis in programmed cell death [2].

(NADH + H+) + CoQ + 4 H+(matrix)->NAD+ + CoQ𝐻2 + 4 H+(intermembrane)

 Figure 1: Showing Complex 1 contains 44 individual polypeptide chains.

 Complex II (Succinate-CoQ Reductase):

Complex II, also known as succinate dehydrogenase, accepts electrons from succinate (an
intermediate in the citric acid cycle) and acts as a second entry point to the ETC. When
succinate oxidizes to fumarate, 2 electrons are accepted by FAD within complex II [3]. FAD
passes them to Fe-S clusters and then to coenzyme Q, similar to complex I. However; no
protons are translocated across the membrane by complex II, therefore less ATP is produced
with this pathway [4].

Succinate + FAD -> Fumarate + 2 H+(matrix) + FAD𝐻2

FAD𝐻2 + CoQ -> FAD + CoQ𝐻2

Glycerol-3-Phosphate dehydrogenase and Acyl-CoA dehydrogenase also accept electrons from

glycerol-3-P and fatty acyl-CoA, respectively . Inclusion of these protein complexes allows for
the donation to the ETC by cytosolic NADH (glycerol-3-P acts as a shuttle to regenerate
cytosolic NAD from NADH) and fatty acids undergoing beta-oxidation within the
mitochondria (acyl-CoA is oxidized to enoyl-CoA in the first step, producing FAD𝐻2 ) [5].

Coenzyme Q, also known as ubiquinone (CoQ), is made up of quinone and a hydrophobic tail.
Its purpose is to function as an electron carrier and transfer electrons to complex III. Coenzyme
Q undergoes reduction to semiquinone (partially reduced, radical form CoQH-) and ubiquinol
(fully reduced CoQ𝐻2 ) through the Q cycle.

CoQ is useful because of its ability to carry and donate electrons and particularly because it
can exist in forms with two extra electrons (fully reduced - ubiquinol), one extra electron (semi-
reduced - ubisemiquinone), or no extra electrons (fully oxidized - ubiquinone). This ability
allows CoQ to provide transition between the first part of the electron transport system that
moves electrons in pairs and the last part of the system that moves electrons one at a time [6].

 Complex III (cytochrome bc1 complex) includes cytochromes

(cyt b, cyt c1) and iron-sulfur clusters.

Complex III, also known as cytochrome c reductase, is made up of cytochrome b, Rieske

subunits (containing two Fe-S clusters), and cytochrome c proteins. A cytochrome is a protein
involved in electron transfer that contains a heme group. The heme groups alternate between
ferrous (𝐹𝑒 2+ ) and ferric (𝐹𝑒 3+) states during the electron transfer [7]. Because cytochrome c
can only accept a single electron at a time, this process occurs in two steps (the Q cycle), in
contrast to the single-step complex I and II pathways. Complex III also releases 4 protons into
the intermembrane space at the end of a full Q cycle, contributing to the gradient. Cytochrome
c then transfers the electrons one at a time to complex IV.

Q Cycle:

In the Q-cycle, electrons are passed from ubiquinol (Q) to cytochrome c using Complex III as
an intermediary docking station for the transfer. Two pair of electrons enter from Q𝐻2 and one
pair is returned to another CoQ𝐻2 to re-make Q𝐻2 . The other pair is donated singly to two
different cytochrome c molecules [8].
 Figure 2: Showing that two pair of electrons enter from Q𝐻2 and one pair is returned

Step 1:

In the Q cycle involves ubiquinol (CoQ) and ubiquinone (CoQ) binding to two separate sites
on complex III. CoQ𝐻2 transfers each electron to a different path. One electron goes to Fe-S
and then cytochrome c, while the second electron is transferred to cytochrome b and then to
CoQ bound at the other site [9]. While this occurs, 2 𝐻 + ions are released into the
intermembrane space, contributing to the proton gradient. CoQ𝐻2 is now oxidized to
ubiquinone and dissociates from the complex. The CoQ bound at the second site enters a
transitional CoQH- radical state from accepting one of the electrons [10].

Step 2:

The second step of the cycle involves a repeat of the first: a new CoQ𝐻2 binds to the first site
and transfers two electrons like before (and 2 more 𝐻+ ions released). Again, one electron
passes to cytochrome c and one to cytochrome b, [11] which this time works to reduce CoQH-
to CoQ𝐻2 before it dissociates from complex III and can be recycled. In this way, one full
cycle appears as follows:

2 CoQ𝐻2 (site 1) + CoQ(site 2) + 2 Cyt c(ox) + 2 H+(matrix) -> 2 CoQ(site 1) +

CoQ𝐻2 (site 2) + 2 Cyt c(red) + 4 H+(intermembrane)
  Complex IV (Cytochrome c Oxidase):

Complex IV contains cytochromes (cyt a and cyta3) and copper centers.

Composition: Electron Flow:

Complex IV, also known as cytochrome c oxidase, oxidizes cytochrome c and transfers the
electrons to oxygen, the final electron carrier in aerobic cellular respiration. The cytochrome
proteins a and a3, in addition to heme and copper groups in complex IV transfer the donated
electrons to the bound dioxygen species, converting it into molecules of water [8]. The free
energy from the electron transfer causes 4 protons to move into the intermembrane space
contributing to the proton gradient. Oxygen reduces via the following reaction:[13][14]

o 2 cytochrome c(red) + ½O2 + 4 H+(matrix) -> 2 cytochrome c(ox) + 1 𝐻2 O +

2 H+(intermembrane)

Flow of Electrons

Flow of Electrons Through the Complexes:

The flow of electrons through the ETC is orchestrated by a series of redox reactions, driving
the pumping of protons across the inner mitochondrial membrane. This creates an
electrochemical gradient that is utilized to generate ATP through oxidative phosphorylation.
Electrons from NADH or FADH2 enter the ETC at Complexes I or II, respectively [12].

Initial Electron Donors:

Complex I (NADH-CoQ Reductase):

 NADH transfers electrons to FMN in Complex I.

 FMN then transfers electrons to a series of iron-sulfur (Fe-S) clusters.
 The electrons are ultimately transferred to ubiquinone (Coenzyme Q), forming
ubiquinol (Q𝐻2 ) [13].

Complex II (Succinate Dehydrogenase):

  FADH2 from the citric acid cycle directly transfers electrons to Complex II.
 Electrons from succinate are transferred to FAD, which then transfers them to the iron-
sulfur clusters in Complex II [14].
 Complex II transfers electrons to ubiquinone, forming ubiquinol (Q𝐻2 ).

Complex III (CoQH2-Cytochrome c Reductase):

 Ubiquinol (QH2) donates electrons to Complex III.

 Electrons are transferred through cytochrome b, cytochrome c1, and the Rieske iron-
sulfur protein.
 Cytochrome c picks up the electrons and shuttles them to Complex IV.

Complex IV (Cytochrome c Oxidase):

 Cytochrome c delivers electrons to Complex IV.
 Electrons are transferred through cytochrome a and cytochrome a3, ultimately reducing
oxygen to form water.

ATP Synthesis and Role of Proton Gradient:

In eukaryotic cells, the vast majority of ATP synthesis occurs in the mitochondria in a process
called oxidative phosphorylation. Even plants, which generate ATP by photophosphorylation
in chloroplasts, contain mitochondria for the synthesis of ATP through oxidative
phosphorylation [15].

Oxidative phosphorylation is linked to a process known as electron transport (Figure 3 ). The

electron transport system, located in the inner mitochondrial membrane, transfers electrons
donated by the reduced electron carriers NADH and FADH2 (obtained from glycolysis, the
citric acid cycle or fatty acid oxidation) through a series of electrons acceptors, to oxygen As
we shall see, movement of electrons through complexes of the electron transport system
essentially “charges” a battery that is used to make ATP in oxidative phosphorylation [16]. In
this way, the oxidation of sugars and fatty acids is coupled to the synthesis of ATP,
effectively.extracting energy from food.
The flow of electrons through the ETC creates a proton gradient across the inner mitochondrial
membrane. This gradient is used by ATP synthase to generate ATP through oxidative
phosphorylation.

Dr. Peter Mitchell introduced a radical proposal in 1961 to explain the mechanism by which
mitochondria make ATP. It is known as the chemiosmotic hypothesis and has been shown over
the years to be correct. Mitchell proposed that synthesis of ATP in mitochondria depends on an
electrochemical gradient, across the mitochondrial inner membrane, that arises ultimately from
the energy of reduced electron carriers, NADH and FADH2 [17].

Further, the proposal states that the gradient is created when NADH and FADH2 transfer their
electrons to an electron transport system (ETS) located in the inner mitochondrial membrane.
Movement of electrons through a series of of electron carriers is coupled to the pumping of
protons out of the mitochondrial matrix across the inner mitochondrial membrane into the space
between the inner and outer membranes. The result is creation of a gradient of protons whose
potential energy can be used to make ATP. Electrons combine with oxygen and protons at the
end of the ETS to make water [18].

ATP synthase:

In oxidative phosphorylation, ATP synthesis is accomplished as a result of protons re-entering

the mitochondrial matrix via the transmembrane ATP synthase complex, which combines ADP
with inorganic phosphate to make ATP. Central to the proper functioning of mitochondria
through this process is the presence of an intact mitochondrial inner membrane impermeable
to protons [17].

When this is the case, tight coupling is said to exist between electron transport and the synthesis
of ATP (called oxidative phosphorylation). Chemicals which permeabilize the inner
mitochondrial membrane to protons cause uncoupling, that is, they allow the protons to leak
back into the mitochondrial matrix, rather than through the ATP synthase, so that the movement
of electrons through the ETS is no longer linked to the synthesis of ATP [19].

Proton Pumping:

As electrons pass through complexes I, III, and IV, there is a release of a small amount of
energy at each step, which is used to pump protons from the mitochondrial matrix (inside of
mitochondrion) and deposit them in the intermembrane space (between the inner and outer
membranes of the mitochondrion). The effect of this redistribution is to increase the electrical
and chemical potential across the membrane [20].

ATP formed in electron transport chain

The protein complex harvesting energy from the proton gradient and using it to make ATP
from ADP is an enzyme that has several names - Complex V, PTAS (Proton Translocating
ATP Synthase), and ATP synthase. Central to its function is the movement of protons through
it (from the intermembrane space back into the matrix) [21]. Protons will only provide energy
to make ATP if their concentration is greater in the intermembrane space than in the matrix
and if ADP is available.

It is possible, in some cases, for the concentration of protons to be greater inside the matrix
than outside of it. When this happens, the ATP synthase can run backwards, with protons
moving from inside to out, accompanied by conversion of ATP to ADP + Pi [22]. This is
usually not a desirable circumstance and there are some controls to reduce its occurrence.

Normally, ATP concentration will be higher inside of the mitochondrion and ADP
concentration be higher outside the mitochondrion. However, when the rate of ATP synthesis
exceeds the rate of ATP usage, then ATP concentrations rise outside the mitochondrion and
ADP concentrations fall everywhere.

This may happen, for example, during periods of rest. It has the overall effect of reducing
transport and thus lowering the concentration of ADP inside the matrix. Reducing ADP
concentration in the matrix reduces oxidative phosphorylation and has effects on respiratory
control [23].

Another important consideration is that when ATP is made in oxidative phosphorylation, it is

released into the mitochondrial matrix, but must be transported into the cytosol to meet the
energy needs of the rest of the cell. This is accomplished by action of the adenine nucleotide
translocase, an antiport that moves ATP out of the matrix in exchange for ADP moving into
the matrix. This transport system is driven by the concentrations of ADP and ATP and ensures
that levels of ADP are maintained within the mitochondrion, permitting continued ATP
synthesis [24].

One last requirement for synthesis of ATP from ADP is that phosphate must also be imported
into the matrix. This is accomplished by action of the phosphate translocase, which is a symport
that moves phosphate into the mitochondrial matrix along with a proton.

There is evidence that the two translocases and ATP synthase may exist in a complex, which
has been dubbed the ATP synthasome.

The electron transport system charges the battery for oxidative phosphorylation by pumping
protons out of the mitochondrion. The intact inner membrane of the mitochondrion keeps the
protons out, except for those that re-enter through ATP Synthase [25]. The ATP Synthase
allows protons to re-enter the mitochondrial matrix and harvests their energy to make ATP.

Conclusion:
The electron transport chain is a highly coordinated process involving multiple protein
complexes embedded within the inner mitochondrial membrane. Through a series of redox
reactions, electrons are transferred, driving the pumping of protons and ultimately leading to
the synthesis of ATP. Understanding the intricacies of the ETC and ATP synthesis is
fundamental to comprehending cellular respiration and energy metabolism.
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