Most of the free energy released during the oxidation of glucose to CO2 is retained in the reduced

coenzymes NADH and FADH2 generated during glycolysis and the citric acid cycle. Electrons are released
from NADH and FADH2 and eventually are transferred to O2 to form H2O through a chain of inner
mitochondrial membrane protein is called electron transport chain or respiratory chain.

The free energy released during oxidation of a single NADH or FADH2 molecule by O2 is sufficient to drive
the synthesis of several molecules of ATP from ADP and Pi, a reaction with a ΔG°′ of +7.3 kcal/mol. The
mitochondrion maximizes the production of ATP by transferring electrons from NADH and FADH 2 through
a series of electron carriers all but one of which are integral components of the inner membrane. This step-
by-step transfer of electrons via the electron transport chain (also known as the respiratory chain) allows
the free energy in NADH and FADH2 to be released in small increments. At several sites during electron
transport from NADH to O2, protons from the mitochondrial matrix are transported uphill across the inner
mitochondrial membrane and a proton concentration gradient forms across it (Figure 16-17). Because the
outer membrane is freely permeable to protons, the pH of the mitochondrial matrix is higher (i.e., the
proton concentration is lower) than that of the cytosol and intermembrane space. An electric potential
across the inner membrane also results from the uphill pumping of positively charged protons outward
from the matrix, which becomes negative with respect to the intermembrane space. Thus free energy
released during the oxidation of NADH or FADH2 is stored both as an electric potential and a proton
concentration gradient — collectively, the proton-motive force — across the inner membrane. The
movement of protons back across the inner membrane, driven by this force, is coupled to the synthesis of
ATP from ADP and Pi by the F0F1 complex.

The synthesis of ATP from ADP and Pi, driven by the transfer of electrons from NADH or FADH2 to O2, is
called oxidative phosphorylation
 Figure : The pathway of electron transport (blue arrows) and proton transport (red arrows) in the inner
mitochondrial membrane

NADH-CoQ Reductase Complex

In the NADH-CoQ reductase complex , electrons first flow from NADH to FMN and then to an iron-sulfur
protein .Iron-sulfur protein donates two electrons to the CoQH2 – cytochrome c reductase complex,
regenerating oxidized CoQ

Four protons are translocated from the mitochondrial matrix across the inner membrane for each pair of
electrons transferred.

Succinate-CoQ Reductase Complex

The two electrons released in conversion of succinate to fumarate are transferred first to FAD, then to an
iron-sulfur carrier, and finally to CoQ, forming the reduced CoQH 2.

No protons are translocated across the membrane by the succinate-CoQ reductase complex

CoQH2—Cytochrome c Reductase Complex

Reduced CoQH2, generated by either oxidation of NADH or succinate, donates two electrons to the
CoQH2 – cytochrome c reductase complex, regenerating oxidized CoQ. Within this complex the released
electrons are transferred to an iron-sulfur protein , then to cytochrome c1. Finally, the two electrons are
transferred to two molecules of the oxidized form of cytochrome c , forming reduced cytochrome c.
Four protons are translocated from the mitochondrial matrix across the inner membrane for each pair of
electrons transferred.

Cytochrome c Oxidase Complex

Cytochrome c, after being reduced by the CoQH2 – cytochrome c reductase complex, transports electrons
to the cytochrome c oxidase complex .Within this complex, electrons are transferred first to a pair of
copper ions (Cua2+), then to cytochrome a, then to a complex of a second copper ion (Cub2+), and
cytochrome a3, and finally to O2, the ultimate electron acceptor, yielding H 2O.

During transport of each pair of electrons through the cytochrome c oxidase complex, two protons are
translocated across the membrane.

Structure of ATP Synthase or F0F1 complex

Ref : Molecular cell biology ,Lodish Ref : Lehninger Principles of Biochemistry

The F0F1 complex, or ATP synthase, has two principal components, F0 and F1. F0 is located within the
membrane and contains a transmembrane channel through which protons flow toward F 1. F0 has multiple
copies of c, one a, and two b subunits .The proton channel lies at the interface of the a and c subunits

The F1 portion is a water-soluble complex of five distinct polypeptides with the composition α3β3γδϵ. Three
α and three β subunits form a hexameric structure that rests atop the single rod-shaped γ subunit, which is
inserted into the c ring of F0. The ϵ subunit is attached to γ and probably also contacts the c subunits of F0.
The δ subunit of the F1 complex contacts the b subunit of the F0 complex;
1. A small protein ,designated HPr ,is phosphorylated by PEP

PEP + HPr Phopsho-HPr + pyruvate

This reaction is not specific to the glucose transport system but is involved in sugar transport of E coli in general.
Enzyme I and HPr serve as catalysts in transport systems for glucose ,mannose ,fructose and other hexoses ; both
proteins are soluble

2. The phosphorylated group of phopsho-HPr is transferred to enzyme III :

Phospho-HPr + enzyme III glc HPr + phosphoenzyme III glc

Enzyme III is substrate specific and the glucose-specific protein is designated enzyme III glc

3. The phosphoryl group of enzyme III glc is transferred to glucose and the glucose-6-phosphate is released into the
cytoplasm .This reaction is catalyzed b enzyme II ,which is specific for glucose and is membrane bound.

phosphoenzyme III glc +glucose glucose-6-phosphate + enzyme III glc

The chemiosmotic theory presumes that :

1. The cytoplasmic membrane is impermeable to OH- and H+

2. The respiratory chain is localized in the membrane in such a way that a PH gradient and a membrane potential
are formed by vectorial extraction and excretion of protons during electron transport

3. The ATP synthesis is so ingeniously constructed that it can take advantage of the PH gradient and the
membrane potential for ATP synthesis from ADP and Pi.