The erythrocyte (red blood cell, RBC) is the

primary blood cell, circulating at 5 million RBCs

per microliter of blood on average. It is anucleate
and biconcave and has an average volume of 90
fL. The cytoplasm provides abundant hemoglobin,
a complex of globin, protoporphyrin, and iron that
transports elemental oxygen (O2) from high
partial pressure to low partial pressure
environments, that is, from lung capillaries to the
capillaries of organs and tissues. Hemoglobin,
plasma proteins, and additional RBC proteins also
transport molecular carbon dioxide (CO2) and
bicarbonate (HCO3−) from the tissues to the
lungs. Hemoglobin is composed of four globin
molecules, each supporting one heme molecule;
each heme molecule contains a molecule of iron.
The biconcave RBC shape supports deformation,
enabling the circulating cell to pass smoothly
through capillaries, where it readily exchanges O2
https://images.app.goo.gl/9QNxbgwv3bVZTdxK8
and CO2 while contacting the vessel wall.
RBCs are produced through erythrocytic (normoblastic) maturation in bone
marrow tissue . The nucleus, while present in maturing marrow normoblasts,
becomes extruded as the cell passes from the bone marrow to peripheral
blood. Cytoplasmic ribosomes and mitochondria disappear 24 to 48 hours
after bone marrow release, eliminating the cells’ ability to produce proteins or
support oxidative metabolism.

Adenosine triphosphate (ATP) is produced within the cytoplasm through

anaerobic glycoglycocalyx. ATP drives mechanisms that slow the
destruction of protein and iron by environmental peroxides and superoxide
anions, maintaining hemoglobin’s function and membrane integrity.
Oxidation, however, eventually takes a toll, limiting the RBC circulating life
span to 120 days, whereupon the cell becomes disassembled into its
reusable components globin, iron, and the phospholipids and proteins of the
cell membrane, while the protoporphyrin ring is excreted as bilirubin.
 Energy production—anaerobic glycolysis

• Lacking mitochondria, the RBC relies on anaerobic

glycolysis for its energy. The hemoglobin exchange of O2
and CO2 is a passive function, however the cells’
metabolic processes require energy. As energy production
slows, the RBC grows senescent and is removed from the
circulation.
Glucose enters the RBC without energy expenditure via the
transmembrane protein Glut-1. Anaerobic glycolysis, the
EMP, requires glucose to generate ATP, a high-energy
phosphate source. With no cytoplasmic glycogen
organelles, RBCs lack internal energy stores and rely on
plasma glucose for glycolysis-generated ATP. Through the
EMP, glucose is catabolized to pyruvate (pyruvic acid),
consuming two molecules of ATP per molecule of glucose
and maximally generating four molecules of ATP per
molecule of glucose, for a net gain of two molecules of ATP.
 First Phase
• The first phase of glycolysis employs glucose
phosphorylation, isomerization, and diphosphorylation to
yield fructose 1,6-bisphosphate (F1,6-BP). Fructose-
bisphosphate aldolase cleaves F1,6-BP to produce
glyceraldehyde-3-phosphate. Intermediate stages
employ, in order, the enzymes hexokinase, glucose-6-
phosphate isomerase, and 6-phosphofructokinase. The
initial hexokinase and 6-phosphofructokinase steps
consume a total of 2 ATP molecules and limit the rate of
glycolysis.
 Glucose Catabolism: First Phase

Substrate Enzymes Products

Glucose, ATP Hexokinase G6P, ADP

G6P Glucose-6-Phosphate F6P

Isomerase
F6P, ATP 6-Phophofructokinase F1,6-BP, ADP

F1,6BP Fructose-biphosphate adolase DHAP, G3P

 Second Phase
The second phase of glucose catabolism converts G3P to
3-phosphoglycerate (3-PG). In the first step, G3P is
oxidized to 1,3-bisphosphoglycerate (1,3-BPG) through the
action of glyceraldehyde-3-phosphate dehydrogenase
(G3PD). 1,3-BPG is dephosphorylated byphosphoglycerate
kinase, which generates 2 ATP molecules and 3-PG.
Substrate Enzymes Products

G3P Glyceride-3-Phosphate 1,3-BPG

dehydrogenase
1,3-BPG, ADP Phosphoglycerate kinase 3PG, ATP

1,3-BPG Biphosphoglycerate mutase 2, 3-BPG

2,3-BPG Biphosphoglycerate Phosphatase 3-PG

 Third Phase
• The third phase of glycolysis converts 3-PG to pyruvate and generates ATP.
Substrates, enzymes, and products are listed in Table 9-3. The product 3-PG
is isomerized byphosphoglycerate mutase to 2-phosphoglycerate (2-PG).
Enolase (phosphopyruvate hydratase) then converts 2-PG to
phosphoenolpyruvate (PEP). Pyruvate kinase (PK) splits off the phosphates,
forming 2 ATP molecules and pyruvate. PK activity is allosterically modulated
by increased concentrations of F1,6-BP, which enhances the affinity of PK for
PEP.5 Thus, when the F1,6-BP is plentiful, increased activity of PK favors
pyruvate production. Pyruvate may diffuse from the erythrocyte or may
become a substrate for lactate dehydrogenase with regeneration of the
oxidized form of nicotinamide adenine dinucleotide (NAD+). The ratio of
NAD+ to the reduced form (NADH) modulates the activity of this enzyme.
Substrate Enzymes Products

3-PG Phosphoglycerate mutase 2-PG

2-PG Enolase (phosphopyruvate PEP

hydratase)

PEP, ADP Pyruvate kinase Pyruvate, ATP

 Glycolysis diversion pathways (shunts)

Three alternate pathways, called diversions or shunts,

branch from the glycolytic pathway. The three diversions
are the hexose monophosphate pathway (HMP) or aerobic
glycolysis, the methemoglobin reductase pathway, and the
Rapoport-Luebering pathway.
 Hexose monophosphate pathway
• Aerobic or oxidative glycolysis occurs through a diversion
of glucose catabolism into the HMP, also known as the
pentose phosphate shunt. The HMP detoxifies peroxide
(H2O2), which arises from O2 reduction in the cell’s
aqueous environment, where it oxidizes and destroys
heme iron, proteins, and lipids, especially lipids containing
thiol groups. By detoxifying peroxide, the HMP extends
the functional life span of the RBC.
• The HMP diverts glucose-6-phosphate (G6P) to ribulose 5-
phosphate by the action of glucose-6-phosphate
dehydrogenase (G6PD). In the process, oxidized nicotinamide
adenine dinucleotide phosphate (NADP) is converted to its
reduced form (NADPH). NADPH is then available to reduce
oxidized glutathione (GSSG) to reduced glutathione (GSH) in
the presence of glutathione reductase. Glutathione is a
cysteine-containing tripeptide, and the designation GSH
highlights the sulfur in the cysteine moiety. Reduced
glutathione becomes oxidized as it reduces peroxide to water
and oxygen via glutathione peroxidase.
• During steady-state glycolysis, 5% to 10% of G6P is
diverted to the HMP. After oxidative challenge, HMP
activity may increase up to thirtyfold. The HMP
catabolizes G6P to ribulose 5-phosphate and carbon
dioxide by oxidizing G6P at carbon 1.
• G6PD provides the only means of generating NADPH for
glutathione reduction, and in its absence erythrocytes are
particularly vulnerable to oxidative damage. With normal
G6PD activity, the HMP detoxifies oxidative compounds
and safeguards hemoglobin, sulfhydryl-containing
enzymes, and membrane thiols, allowing RBCs to safely
carry O2. However, in G6PD deficiency, the most
common inherited RBC enzyme deficiency worldwide, the
ability to detoxify is hampered, resulting in hereditary
nonspherocytic anemia.
 Methemoglobin reductase pathway
• Heme iron is constantly exposed to oxygen and peroxide. Peroxide
oxidizes heme iron from the ferrous (+2) to the ferric (+3) state. The
affected hemoglobin molecule is called methemoglobin. Although the
HMP prevents hemoglobin oxidation by reducing peroxide, it is not able to
reduce methemoglobin once it forms. NADPH is able to do so, but only
slowly. The reduction of methemoglobin by NADPH is rendered more
efficient in the presence of methemoglobin reductase, also called
cytochrome b5 reductase. Using H+ from NADH formed when G3P is
converted to 1,3-BPG, cytochrome b5 reductase acts as an intermediate
electron carrier, returning the oxidized ferric iron to its ferrous, oxygen-
carrying state. This enzyme accounts for more than 65% of the
methemoglobin-reducing capacity within the RBC.
 Rapoport-luebering pathway
• A third metabolic shunt generates 2,3-
bisphosphoglycerate (2,3-BPG; also called 2,3-
diphosphoglycerate or 2,3-DPG). 1,3-BPG is diverted by
bisphosphoglycerate mutase to form 2,3-BPG. 2,3-BPG
regulates oxygen delivery to tissues by competing with
oxygen for the oxygen-binding site of hemoglobin. When
2,3-BPG binds heme, oxygen is released, which
enhances delivery of oxygen to the tissues.
• 2,3-BPG forms 3-PG by the action of bisphosphoglycerate
phosphatase. This diversion of 1,3-BPG to form 2,3-BPG
sacrifices the production of two ATP molecules. There is further
loss of two ATP molecules at the level of PK, because fewer
molecules of PEP are formed. Because two ATP molecules were
used to generate 1,3-BPG and production of 2,3-BPG eliminates
the production of four molecules, the cell is put into ATP deficit by
this diversion. There is a delicate balance between ATP
generation to support the energy requirements of cell metabolism
and the need to maintain the appropriate oxygenation and
deoxygenation status of hemoglobin.
• Acidic pH and low concentrations of 3-PG and 2-PG
inhibit the activity of bisphosphoglycerate mutase, thus
inhibiting the shunt and retaining 1,3-BPG in the EMP.
These conditions and decreased ATP activate
bisphosphoglycerate phosphatase, which returns 2,3-
BPG to the glycolysis mainstream. In summary, these
conditions favor generation of ATP by causing the
conversion of 1,3-BPG directly to 3-PG and returning 2,3-
BPG to 3-PG for ATP generation downstream by PK.