Embden-Meyerhof Pathway

Embden-Meyerhof pathway of Glycolysis is the anaerobic metabolic pathway

involving 10 steps, where
glucose C6H12O6 is converted
into pyruvate, CH3COCOO− + H+. The
glycolytic pathway can be divided into
two phases; first phase is the
preparatory phase, also known as
glucose activation phase and the
second phase is the pay-off phase also
called energy extraction phase. During
the first phase, two molecules of ATP
are invested and the hexose chain is
cleaved into two triose phosphates. In
this phase, at first, Glucose is
phosphorylated to form glucose-6-
phosphate. The reaction is catalyzed
by the key glycolytic enzyme,
hexokinase. This enzyme splits the
ATP into ADP, and the Pi (phosphate)
is added onto the glucose. This step is
a regulatory step in glycolysis, hence
it is irreversible. Next, Glucose-6-
phosphate is isomerized to fructose-6-
phosphate by the
enzyme phosphohexose isomerase. Fructose-6-phosphate is then further
phosphorylated to fructose 1,6-bisphosphate by the enzyme
phosphofructokinase-1. This enzyme also catalyzes the transfer of a
phosphate group from ATP to fructose-6-phosphate and the reaction is
irreversible. Then, the 6-carbon fructose-1,6-bisphosphate is cleaved into two
3 carbon units; one glyceraldehyde-3-phosphate (GAP) and one molecule of
dihydroxy acetone phosphate (DHAP). GAP is on the direct pathway of
glycolysis, whereas DHAP is not. Hence, the enzyme Triose-phosphate
isomerase converts DHAP into GAP which is useful for generating ATP. As a
net result, glucose is now cleaved into 2 molecules of glyceraldehyde-3-
phosphate. The first phase does not generate energy, but two ATP has been
used in this phase.
In payoff phase, oxidation of glucose releases energy in the form of ATP and
NADH.  Starting the pay-off phase, glyceraldehyde 3-phosphate is oxidized to
1,3-bisphosphoglycerate catalyzed by the enzyme glyceraldehyde 3-
phosphate dehydrogenase. In this step, NAD+ is reduced to NADH and this
reaction is an energy yielding reaction. Next, ATP and 3-phosphoglycerate is
formed due to the transferring of the phosphoryl group from the carboxyl
group of 1,3-bisphosphoglycerate to ADP, by the enzyme phosphoglycerate
kinase. Then, the 3-phosphoglycerate is isomerized to 2-phosphoglycerate
by shifting the phosphate group from 3rd carbon atom to 2nd carbon atom
by the enzyme phosphoglucomutase. Afterwards, 2-phosphoglycerate is
converted to phosphoenolpyruvate (PEP) by the enzyme enolase and this
reaction involves the removal of a water molecule. Lastly,
phosphoenolpyruvate (PEP) is dephosphorylated to pyruvate, by pyruvate
kinase which is a key glycolytic enzyme. Hence this step is irreversible.

During this phase, since Glucose splits to give two molecules of

Glyceraldehyde-3-phosphate, each step in this phase occurs twice per
molecule of glucose and coupled formation of ATP take place. The net energy
yield of this reaction is 2 NADH and 2 ATP. Additionally, 2 Pyruvate is also
formed.

Functions of Embden-Meyerhof Pathway

Glycolytic breakdown of glucose is the sole source of metabolic energy in the
red blood cells. Red blood cells cannot depend on aerobic glycolysis, to
extract energy from glucose. Therefore, they use the Embden-Meyerhof
pathway to anaerobically process glucose into usable energy, or adenosine
triphosphate (ATP). This ATP is necessary for active transport of cations
across the red cell membrane, hence maintaining the activity of sodium-
potassium pump, reducing the occurrence of unfavorable osmotic effects
such as swelling of the cell and regulating cellular volume. The ATP is also
used for the restoration and preservation of membrane integrity as well as
maintenance of an active membrane barrier against efflux of small
molecules and ions from cell.
Apart from ATP production, the Embden Meyerhof pathway maintains
pyridine nucleotides in a reduced state. The NADH that is generated from the
reduction of NAD+ in Embden Meyerhof pathway is an important contributor
to the enzyme methemoglobin reductase. This enzyme assists in the
reduction of methemoglobin to haemoglobin and maintain iron in its normal
ferrous state. NADH acts as the electron donor in this process thus,
highlighting its importance in the metabolic pathway.
Moreover, Pyruvate is a precursor molecule for lactic acid fermentation which
maintains NAD+ concentration. This NAD+ is essential for the formation of
2,3-diphosphoglycerate (2,3-DPG), which is an important regulator of the
oxygen-carrying capacity of red blood cells. Apart from modulating
haemoglobin oxygen affinity, 2,3-DPG constitutes an energy buffer.
Furthermore, pyruvate produced here is converted to acetyl-CoA and used in
the Kreb’s cycle. Additionally, most of the reactions of this glycolytic pathway
are reversible, which are also used for gluconeogenesis.
Effect of impairments in the pathway
Impairments in this Embden Meyerhof pathway leads to occurrence of
several clinical and hematologic changes. While glycolytic enzymopathies
are the most common, other impairments also occur. Looking into the
impairments, defects in the Luebering-Rapaport bypass (a branch of the
normal glycolytic pathway, producing 2,3-diphosphoglycerate (2,3-DPG), can
affect the levels of 2,3-DPG available to erythrocytes, leading to hindering of
the regulation of the oxygen-carrying capacity of red blood cells.
The red blood cells have to preserve cations intracellularly and active
transport of these molecules require ATP generated from the Embden
Meyerhof pathway. Impairments in this pathway cause less ATP to be
produced leading to the cells being unable to energize the metabolic pumps,
which further causes the cells to have uncontrolled cation flux and the cell
will not survive normally. An example of such is the Pyruvate kinase
deficiency, where red blood cells cannot produce sufficient amount of the
enzyme pyruvate kinase which is crucial for the ATP production, leading to
leakage of potassium and water in the cell while the calcium ion
concentrations increase. This causes cell to becoming rigid, lose flexibility,
and susceptibility of cell to early splenic sequestration and eventual
hemolysis. On the other hand, elevation of this enzyme levels has been
identified as a source of polycythemia, the elevation of hematocrit.
Moreover, increases in the amount of cellular Pyruvate Kinase also result in
decreases in 2,3-DPG concentrations. This triggers tissue hypoxia causing
reluctancy of hemoglobin to release oxygen. Another enzyme defect is
Triosephosphate isomerase (TPI) deficiency. TPI is the glycolytic enzyme
which catalyzes the interconversion of glyceraldehyde-3-phosphate and
DHAP. A deficiency of TPI causes hereditary nonspherocytic hemolytic
anemia, progressive neurologic dysfunction and increased susceptibility to
infection. Furthermore, Glucose 6-phosphate isomerase (GPI) catalyzes the
interconversion of G6P into fructose-6-phosphate (F6P) in the second step of
the Embden-Meyerhof pathway. GPI deficiency causes hemolytic anemia of
variable severity and in rare cases, it also affects nonerythroid tissues,
causing neurologic symptoms and granulocyte dysfunction.