Electron Transport and

Oxidative Phosphorylation
• Energy obtained by Substrate level
phosphorylation in glycolysis in the form of
ATP
• But mostly via oxidation–reduction reactions
into NADH and reduced flavoproteins, the
latter symbolized by [FADH2].
Where in the Cell Are ElectronTransport and
Oxidative Phosphorylation Carried Out?
• oxidative phosphorylation are membrane
• associated.
• In bacteria it is in cytoplasmic membrane
• In eukaryotes it is in Mitochondria, also here
fatty acid oxidation and TCA cycle
• Mitochondria number in cells varies
• RBCs have no Mitochondria
• Mitochondria have Outer membrane:The smooth outer membrane is
about 30% to 40% lipid and 60% to 70% protein and has a relatively
high concentration of phosphatidylinositol. significant amounts of porin
• Inner membrane: richly packed with proteins, which account for nearly
80% of its weight, The fatty acids of inner membrane lipids are highly
unsaturated. Cardiolipin and diphosphatidylglycerol are abundant.
extensively folded called cristae.
• Inter membrane space.
• The space inside the inner mitochondrial membrane is called the
matrix, and it contains most of the enzymes of the TCA cycle and fatty
acid oxidation.
• Mitochondria contain circular DNA, ribosomes and DNA synthesis
enzymes
 The Electron-Transport Chain Can Be
Isolated in Four Complexes
• Flavoproteins, which contain tightly bound FMN or FAD as prosthetic
groups and which may participate in one- or two electron transfer
events.
• Coenzyme Q, also called ubiquinone (and abbreviated CoQ or UQ)
which can function in either one- or two-electron transfer reactions.
• Several cytochromes (proteins containing heme prosthetic groups
which function by carrying or transferring electrons), including
cytochromes b, c, c 1, a, and a3. Cytochromes are one-electron transfer
agents in which the heme iron is converted from Fe2 to Fe3 and back.
• A number of iron–sulfur proteins, which participate in one-electron
transfers involving the Fe2 and Fe3 states.
• Protein-bound copper, a one-electron transfer site that converts
between Cu+ and Cu2.
• The components of the electron-transport chain can be purified from the
mitochondrial inner membrane. Solubilization of the membranes containing the
electron-transport chain results in the isolation of four distinct protein complexes, and
the complete chain can thus be considered to be composed of four parts:
• (I) NADH–coenzyme Q reductase,
• (II) succinate–coenzyme Q reductase,
• (III) coenzyme Q–cytochrome c reductase, and
• (IV) cytochrome c oxidase
• Complex I accepts electrons from NADH, serving as a link between glycolysis, the TCA
cycle, fatty acid oxidation, and the electron-transport chain.
• Complex II includes succinate dehydrogenase and thus forms a direct link between the
TCA cycle and electron transport. Complexes I and II produce a common product,
reduced coenzyme Q (UQH2),
• Complex III oxidizes UQH2 while reducing cytochrome c, which in turn is the substrate
for Complex
• IV, cytochrome c oxidase. Complex IV is responsible for reducing molecular oxygen.
 Complex I Oxidizes NADH and Reduces
Coenzyme Q
• This complex transfers a pair of electrons from
NADH to coenzyme Q,
• This enzyme complex is NADH dehydrogenase
• It contains more than 30 polypeptide chains, 1
molecule of flavin mononucleotide (FMN), and
as many as seven Fe-S clusters
• By virtue of its dependence on FMN, NADH–
UQ reductase is a flavoprotein.
Mechanism of the NADH–UQ reductase
• Although unknown completely
• First step: Involves binding of NADH to the
enzyme on the matrix side of the inner
mitochondrial membrane and transfer of
electrons from NADH to tightly bound FMN:
• The second step: Involves the transfer of electrons
from the reduced [FMNH2] to a series of Fe-S
proteins.
• The final step of the reaction: Involves the transfer of
two electrons from iron–sulfur clusters to coenzyme Q
• Coenzyme Q is a mobile electron carrier. Its
isoprenoid tail makes it highly hydrophobic, and it
diffuses freely in the hydrophobic core of the inner
mitochondrial membrane. As a result, it shuttles
electrons from Complexes I and II to Complex III.
 Complex I Transports Protons from the
Matrix to the Cytosol
• The oxidation of one NADH and the reduction of one
UQ by NADH–UQ reductase results in the net
transport of protons from the matrix side to the
cytosolic side of the inner membrane.
• The cytosolic side, where H accumulates, is referred
to as the P (for positive) face; similarly, the matrix
side is the N (for negative) face.
• Some of the energy liberated by the flow of electrons
through this complex is used in a coupled process to
drive the transport of protons across the membrane.
Complex II Oxidizes Succinate and Reduces
Coenzyme Q
• Complex II----------succinate dehydrogenase, the only
TCA cycle enzyme that is an integral membrane
protein in the inner mitochondrial membrane.
• It contains Fe-S centres, Ppolypeptides, FAD.
• When succinate is converted to fumarate in the TCA
cycle, concomitant reduction of bound FAD to FADH2
occurs in succinate dehydrogenase.
• FADH2 transfers its electrons immediately to Fe-S
centers, which pass them on to UQ. Electron flow
from succinate to UQ.
Oxidation of one FADH2 in the electron-transport chain results in
synthesis of approximately two molecules of ATP, compared with the approximately
three ATPs produced by the oxidation of one NADH.
Complex III Mediates Electron Transport from Coenzyme Q
to Cytochrome c

• In the third complex of the electron-transport

chain, reduced coenzyme Q (UQH2) passes its
electrons to cytochrome c via a unique redox
pathway known as the Q cycle.
• UQ–cytochrome c reductase (UQ–cyt c
reductase).
• Three different cytochromes and an Fe-S
protein
• In the cytochromes of these and similar
complexes, the iron atom at the center of the
porphyrin ring cycles between the reduced
Fe2 (ferrous) and oxidized Fe3 (ferric) states.
• Cytochromes were first named and classified on the
basis of their absorption spectra
• The b cytochromes contain iron protoporphyrin IX
(Figure 20.10),the same heme found in hemoglobin
and myoglobin.
• The c cytochromes contain heme c, derived from
iron protoporphyrin IX
• UQ–cyt c reductase contains a b –type cytochrome,
of 30 to 40 kD, with two different heme sites and
one c -type cytochrome.
• UQ–cyt c reductase, also known as the cytochrome bc 1
complex,
• The complex is a dimer, with each monomer consisting of 11
protein subunits and 2165 amino acid residues (monomer mass,
248 kD). The dimeric structure is pear-shaped and consists of a
large domain that extends 75 Å into the mitochondrial matrix, a
transmembrane domain consisting of 13 transmembrane α-
helices in each monomer and a small domain that extends 38 Å
into the intermembrane space.
• Most of the Rieske protein (an Fe-S protein named for its
discoverer) is mobile in the crystal. mobility of this subunit could
be required for electron transfer in the function of this complex.
 Q cycle
• A large pool of UQ and UQH2 exists in the inner
mitochondrial membrane. The Q cycle is initiated when a
molecule of UQH2 from this pool diffuses to a site (called
Qp) on Complex III near the cytosolic face of the membrane.
• Oxidation of this UQH2 occurs in two steps.
• First, an electron from UQH2 is transferred to the Rieske
protein and then to cytochrome c1.
• This releases two H+ to the cytosol and leaves UQ.- , a
semiquinone anion form of UQ, at the Qp site.
• The second electron is then transferred to the bL heme,
converting UQ.- to UQ.
• The electron on the bL heme facing the cytosolic
side of the membrane is now passed to the bH
heme on the matrix side of the membrane
• The electron is then passed from bH to a
molecule of UQ at a second quinone-binding site,
Qn, converting this UQ to UQ-. . The resulting
UQ-. remains firmly bound to the Qn site. This
completes the first half of the Q cycle
• The second half of the cycle is similar to the first half,
with a second molecule of UQH2 oxidized at the Q p
site, one electron being passed to cytochrome c1 and
the other transferred to heme bL and then to heme bH.
• In this latter half of the Q cycle, however, the bH
electron is transferred to the semiquinone anion, UQ-. ,
at the Qn site. With the addition of two H+ from the
mitochondrial matrix, this produces a molecule of
UQH2, which is released from the Qn site and returns
to the coenzyme Q pool, completing the Q cycle.
 Cytochrome c Is a Mobile Electron Carrier

• Electrons traversing Complex III are passed through

cytochrome c1 to cytochrome c. Cytochrome c is the only
one of the cytochromes that is water soluble.
• Cytochrome c, like UQ, is a mobile electron carrier. It
associates loosely with the inner mitochondrial
membrane (in the intermembrane space on the cytosolic
side of the inner membrane) to acquire electrons from
the Fe-S–cyt c 1 aggregate of Complex III, and then it
migrates along the membrane surface in the reduced
state, carrying electrons to cytochrome c oxidase, the
fourth complex of the electron-transport chain.
Complex IV Transfers Electrons from Cytochrome c to Reduce
Oxygen on the Matrix Side

• Complex IV is called cytochrome c oxidase because it accepts

electrons from cytochrome c and directs them to the four-
electron reduction of O2 to form H2O: (nxt page plz)
• Thus, O2 and cytochrome c oxidase are the final destination
for the electrons derived from the oxidation of food materials.
In concert with this process, cytochrome c oxidase also drives
transport of protons across the inner mitochondrial
membrane.
• These important functions are carried out by a
transmembrane protein complex consisting of more than ten
subunits.
 Electron Transfer in Complex IV Involves
Two Hemes and Two Copper Sites
• Cytochrome c oxidase contains two heme centers
(cytochromes a and a3) as well as two copper atoms.
The copper sites, CuA and CuB, are associated with
cytochromes a and a3, respectively.
• The copper sites participate in electron transfer by
cycling between the reduced (cuprous) Cu state and the
oxidized (cupric) Cu2 state. (Remember, the
cytochromes and copper sites are oneelectron transfer
agents.) Reduction of one oxygen molecule requires
passage of four electrons through these carriers—one
at a time.
• Electrons from cytochrome c are transferred to CuA sites and
then passed to the heme iron of cytochrome a. CuA is liganded
by two cysteines and two histidines.
• The heme of cytochrome a is liganded by imidazole rings of
histidine residues.
• The CuA and the Fe of cytochrome a are within 1.5 nm of each
other.
• CuB and the iron atom of cytochrome a3 are also situated close
to each other and are thought to share a ligand, which may be a
cysteine sulfur.
• This closely associated pair of metal ions is referred to as a
binuclear center.
• The electron pathway through Complex IV continues as CuB accepts a
single electron from cytochrome a (state O→state H).
• A secondelectron then reduces the iron center to Fe2 (H→R), leading to
the bindingof O2 (R→A) and the formation of a peroxy bridge between
heme a3 and CuB (A→P). This amounts to the transfer of two electrons
from the binuclear center to the bound O2.
• The next step involves uptake of two H and a third electron (P→F), which
leads to cleavage of the O-O bond and generation of an unusual Fe4 state
at the heme.
• Uptake of a fourth e facilitates formation of ferric hydroxide at the heme
center (F→O’).
• In the final step of the cycle (O’→O), protons from the mitochondrial
matrix are accepted by the coordinated hydroxyl groups, and the resulting
water molecules dissociate from the binuclear center.