ERYTHROCYTE METABOLISM

AND MEMBRANE STRUCTURE

AND FUNCTION
Chapter 9
 Erythrocyte (Red Blood Cell)
◦ 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 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 and CO2 while contacting the vessel wall.
◦ RBCs are produced through erythrocytic (normoblastic)
maturation in bone marrow tissue.
◦ 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 glycolysis (Embden-Meyerhof pathway, EMP) for the lifetime
of the cell.
◦ ATP mechanisms:
• slow the destruction of protein and iron by environmental peroxides and superoxide
anions.
• maintaining hemoglobin’s function and membrane integrity.

oRBC 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
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 listed require energy.
Erythrocyte Metabolic Processes Requiring Energy:
◦ Intracellular cationic gradient maintenance
◦ Maintenance of membrane phospholipid distribution
◦ Maintenance of skeletal protein deformability
◦ Maintenance of functional hemoglobin with ferrous iron
◦ Protecting cell proteins from oxidative denaturation
◦ Glycolysis initiation and maintenance
◦ Glutathione synthesis
◦ Nucleotide salvage reactions
oAs energy production slows, the RBC grows senescent and is
removed from the circulation.

oHematologists have identified hereditary deficiencies of nearly

every glycolytic enzyme, and their common result is shortened
RBC survival, known collectively as hereditary nonspherocytic
hemolytic anemia.
Embden-Meyerhof Pathway
(Anaerobic Pathway of Glucose
Metabolism)
Generates 90- 95% of energy needed by RBC’s
Glucose enters the RBC without energy expenditure via the transmembrane
protein Glut-1.
the EMP, requires glucose to generate 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.
Functions in the maintenance of RBC shape and flexibility
Glucose Catabolism: 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 (G3P)
◦ Intermediate stages employ, in order, the enzymes:
Hexokinase
glucose-6-phosphate isomerase
6-phosphofructokinase

◦ The initial hexokinase and 6-phosphofructokinase steps consume a total of 2 ATP

molecules and limit the rate of glycolysis.
 Glucose Catabolism: 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 by phosphoglycerate kinase, which
generates 2 ATP molecules and 3-PG.
 Glucose Catabolism: Third Phase
◦ The third phase of glycolysis converts 3-PG to pyruvate and generates ATP.
◦ The product 3-PG is isomerized by phosphoglycerate 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. 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.
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
◦ Methemoglobin reductase pathway
◦ Rapport-Luebering pathway
Hexose monophosphate Pathway
◦ Detoxifies peroxide, which arises from o2 reduction in the cell’s aqueous environment,
where it oxidizes and destroys heme iron, protein and lipids, especially lipids containing
thiol groups. By detoxifying peroxide, the HMP extends the functional life span of the RBC.
◦ G6PD provides the only means of generating NADPH For glutathione reduction, and in its
absence erythrocytes are particularly vulnerable to oxidative damage.
◦ NADPH is end product
◦ NADPH is then available to reduce oxidized glutathione (GSSG) to reduced glutathione
(GSH) in the presence of glutathione reductase.
◦ Reduced glutathione becomes oxidized as it reduces peroxide to water and oxygen via
glutathione peroxidase.
◦ Protects the RBC from oxidative injury.
◦ Most common defect is deficiency of the enzyme glucose-6-phosphate dehydrogenase
(G-6PD).
 Methemoglobin Reductase pathway
◦ Maintains iron in the ferrous (Fe++) state.
◦ NADPH is able to reduce methemoglobin but only slowly.
◦ The reduction of methemoglobin by NADPH is rendered more
efficient in the presence of methemoglobin reductase called
cytochrome b5 reductase.
◦ Cytochrome b5 reductase acts as an intermediate electron
carrier, returning the oxidized ferric iron to its ferrous, oxygen
carrying state.
◦ In the absence of the enzyme (methemoglobin reductase),
methemoglobin accumulates and it cannot carry oxygen.
Rapoport –Leubering Shunt
◦ Allows the RBC to regulate oxygen transport during conditions of
hypoxia or acid-base imbalance.
◦ Permits the accumulation of 2,3-DPG which is essential for
maintaining normal oxygen tension, regulating hemoglobin
affinity.
◦ When 2,3-BPG binds heme, oxygen is released which enhances
delivery of oxygen to the tissues.
The Pathway
There are 2 steps in this shunt:
1. Bisphosphoglycerate mutase
converts 1,3-BPG in to 2,3-BPG.
2. 2,3-BPG is hydrolysed to 3-
phosphoglycerate by 2,3-
bisphosphoglycerate
phosphatase.
ATP Yield:
Mature RBCs contain no mitochondria, thus
they depend only upon glycolysis for energy
production 2 ATP
RBC Membrane Deformability
RBC’s are biconcave and average 90 fL in volume.
Average surface area: 140 µm². a 40% excess of
surface area compared with a 90 Fl sphere.
This excess surface to volume ratio enables RBC’s to
stretch undamaged up to 2.5 times their resting
diameter as they pass through narrow capillaries and
through splenic pores 2µm in diameter; this property is
called RBC deformability.
RBC deformability depends not only on RBC geometry but
also on relative cytoplasmic (hemoglobin) viscosity.

Normal mean cell hemoglobin concentration (MCHC) ranges

from 32% to 36%, and as MCHC rises, internal viscosity rises.

As RBC’s age, they lose membrane surface area, while

retaining hemoglobin.

As the MCHC rises, the RBC, unable to pass through the
splenic pores, is destroyed by splenic macrophages.
RBC Membrane Lipids
RBC membrane consist: approximately 8% carbohydrates, 52%
proteins, and 40% lipids.
Phospholipids form an impenetrable fluid barrier as their
hydrophilic polar head groups are arrayed upon the
membrane’s surfaces, oriented both the aqueous plasma and
the cytoplasm.
Hydrophobic nonpolar acyl tails arrange themselves to form a
central layer dynamically sequestered (hidden) from the
aqueous plasma and cytoplasm.
◦ Cholesterol, esterified and largely hydrophobic, resides parallel to the
acyl tails of the phospholipids, equally distributed between the outer
and inner layers, and evenly dispersed within each layer, approximately
one cholesterol molecule per phospholipid molecule.

◦ Cholesterol’s B-hydroxyl group is the only hydrophilic portion, while the

rest of the molecules becomes intercalated among and parallel to the
acyl tails.

◦ The ratio of cholesterol to phospholipid remains relatively constant and

balances the need for deformability and strength.
◦ Membrane enzymes maintains the cholesterol concentration by
regularly exchanging membrane and plasma cholesterol.
◦ Deficiencies in these enzymes are associated with membrane
abnormalities such as acanthocytosis.

◦ Phospholipids are asymmetrically distributed. Phosphatidylcholine and

sphingomyelin predominate in the outer layer; phosphatidyldserine and
phosphatidylthanolamine form most of the inner layer.

◦ Membrane associated enzymes termed flippases, floppases and

scramblases.
◦ Membrane phospholipids and cholesterol may also redistribute laterally so that the
RBC membrane may respond to stresses and deform within 100 milliseconds of being
challenged by the presence of the narrow passage, such as when arriving an
capillary.

◦ Glycolipids (sugar-bearing lipids) make up 5% of external half of the RBC membrane.

They associate in clumps or rafts and support carbohydrate side chains that extend
into the aqueous plasma to anchor the glycocalyx (prevents microbial attack and
protects the RBC from mechanical damage caused by adhesion to neighboring
RBC’s or to the endothelium.
RBC Membrane Proteins
Transmembrane (integral)
Cytoskeletal (skeletal, peripheral) proteins

make up 52% of the membrane structure by mass

At least 300 RBC membrane proteins including 105
transmembrane proteins.

50 have been characterized and named. (ex. receptor

protein, channel protein, etc.)
Transmembrane Proteins
◦ Are a class of integral proteins that penetrate into or
through the membrane bilayer.

◦ They serve as transport and adhesion sites and signaling

receptors .
 Disruption in transport protein

Rise in viscosity and loss of deformability

 Any change affecting adhesion proteins

fragmentation, reducing membrane flexibility,

and shortening the RBC life span
BLOOD GROUP ANTIGEN
◦-defined as a transmembrane proteins
support carbohydrate.
◦ Support the majority of ABH system carbohydrate
determinants by virtue of their copy numbers:
1. Band 3 (anion transport)
2. Glut-1 (glucose transport)
Transmembrane or integral
membrane proteins
◦Glycophorin A- carries the peptide defined M
and N determinants

◦Glycophorin B- carries the Ss determinants which

together comprise the MNSs system.
◦RH system employs two multipass
transmembrane lipoproteins and a
multipass glycoprotein, each of which
crosses the membrane 12 times.
◦Loss of the RhAG glycoprotein prevents
expression of both the D ad CcEe antigens.