Erythrocyte Metabolism and

Membrane
Structure and Function
Chapter 6
Dr. Usamah Sayed

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 ENERGY PRODUCTION—ANAEROBIC GLYCOLYSIS
• Lacking mitochondria, the RBC relies on anaerobic glycolysis for its energy.

• The cells’ metabolic processes require energy. As energy production slows, the RBC grows

senescent and is removed from circulation.

• Anaerobic glycolysis, or Embden-Meyerhof pathway (EMP), requires glucose to generate ATP.

• RBCs lack internal energy stores and rely on plasma glucose to enter the cell to generate ATP.

• Glucose enters the RBC through facilitated diffusion via the transmembrane protein Glut-1.

• Glucose is then catabolized to pyruvate (pyruvic acid) in the EMP, generating four molecules of

ATP per molecule of glucose, for a net gain of two molecules of ATP.

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• The first phase of glycolysis employs glucose phosphorylation, isomerization, and diphosphorylation to
yield fructose 1,6-bisphosphate (F1,6-BP).

• The initial hexokinase and 6-phosphofructokinase steps consume a total of two ATP molecules and limit
the rate of glycolysis.

• Fructose-bisphosphate aldolase then cleaves F1,6-BP to produce glyceraldehyde-3-phosphate (G3P).

• The second phase of glucose catabolism converts G3P to 3-phosphoglycerate (3-PG).

• G3P is oxidized to 1,3-bisphosphoglycerate (1,3-BPG) through the action of glyceraldehyde-3-phosphate

dehydrogenase (G3PD) with the reduction of NAD to NADH.

• 1,3-BPG is dephosphorylated by phosphoglycerate kinase, which generates two ATP molecules and 3-PG.

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• The third phase of glycolysis converts 3-PG to pyruvate and generates ATP.

• The 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 two ATP molecules and pyruvate.

• Pyruvate may diffuse from the erythrocyte or may become a substrate for lactate dehydrogenase (LD
or LDH) with regeneration of the oxidized form of nicotinamide adenine dinucleotide (NAD 1).

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GLYCOLYSIS DIVERSION PATHWAYS (SHUNTS)
Three alternate pathways, called diversions or shunts, branch from the
glycolytic pathway.
The three diversions are:
1. The hexose monophosphate pathway (HMP),
2. The methemoglobin reductase pathway, and
3. The Rapoport-Luebering pathway.

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 RBC MEMBRANE
RBC Membrane Deformability

• RBCs are biconcave, 7 to 8 µm in diameter, with an MCV range of 80 to 100 fL and a mean volume of 90 fL.

• Their average surface area is 140 µm2, which is a 40% excess of surface area compared with a sphere of 7

to 8 µm in diameter.

• This excess surface area-to-volume ratio enables RBCs to stretch undamaged up to 2.5 times their resting

diameter as they pass through narrow capillaries and splenic pores 2 µm in diameter. This property is

called RBC deformability.

• The RBC plasma membrane is 100 times more elastic than a comparable latex membrane, yet it has tensile

(lateral) strength greater than that of steel.

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• The deformable RBC membrane provides the broad surface area and close tissue contact necessary to support

the delivery of O2 from the lungs to body tissues and to transport CO2 from body tissues to the lungs.

• RBC deformability depends not only on RBC geometry but also on relative cytoplasmic (hemoglobin) viscosity.

• The normal mean cell hemoglobin concentration (MCHC) ranges from 32% to 36%, and as MCHC rises,

internal viscosity rises.

• As RBCs age, they lose membrane surface area, while retaining hemoglobin.

• As the MCHC rises, the RBC, unable to pass through the splenic pores, is phagocytized and destroyed by

splenic macrophages.

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RBC Membrane Lipids

• Besides geometry and viscosity, membrane elasticity (pliancy) also

contributes to deformability.

• The RBC membrane consists of approximately 8% carbohydrates,

52% proteins, and 40% lipids.

• The lipid portion, equal parts of cholesterol and phospholipids, forms a bilayer universal to all animal cells.

• Phospholipids form an impenetrable fluid barrier as their hydrophilic polar head groups are arrayed on the

membrane’s surfaces, oriented toward both the aqueous plasma and the cytoplasm, respectively, as

depicted in the fluid mosaic membrane model (FMMM).

• The phospholipids provide a dynamic fluidity to the membrane; if a portion of the lipid bilayer is lost, the

membrane can self-seal to retain the cytoplasmic contents.

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• The membrane also maintains extreme differences in osmotic pressure,

cation concentrations, and gas concentrations between external plasma and

the cytoplasm through the dynamic interaction of the lipids and proteins.

• Cholesterol, esterified and largely hydrophobic, resides parallel to the acyl

tails of the phospholipids.

• It is equally distributed between the outer and inner layers of the phospholipid bilayer, and evenly dispersed

within each layer, with approximately one cholesterol molecule per phospholipid molecule.

• The b-hydroxyl group of cholesterol, the only hydrophilic domain of the molecule, anchors within the

phospholipid polar heads, while the rest of the molecule becomes intercalated among and parallel to the

acyl tails.

• Cholesterol confers tensile strength to the lipid bilayer.

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• As cholesterol concentration rises, the membrane gains strength but loses elasticity.

• The ratio of cholesterol to phospholipids remains relatively constant to maintain the balance of

deformability or elasticity and strength.

• Membrane enzymes maintain the cholesterol concentration by regularly exchanging membrane and

plasma cholesterol.

• The phospholipids are asymmetrically distributed.

• Phosphatidylcholine and sphingomyelin predominate in the outer layer; phosphatidylserine (PS) and

phosphatidylethanolamine form most of the inner layer. Distribution of these four phospholipids is

energy dependent, relying on a number of membrane-associated enzymes.

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• When phospholipid distribution is disrupted, PS, the only negatively charged phospholipid,

redistributes to the outer layer. Splenic macrophages possess receptors that bind to the PS

displayed on senescent and damaged RBCs and remove them from circulation.

• 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 a narrow passage, such as a capillary.

• As the proportion of cholesterol increases, however, the RBC becomes more rigid and is

unable to deform as readily.

• As a result, the cell membrane surface area-to-volume ratio increases giving the RBC a

target cell appearance.

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• Glycolipids (sugar-bearing lipids) make up 5% of the external half of the RBC membrane (Figure 6.2).

• They associate in clumps or rafts and support carbohydrate side chains that extend into the aqueous

plasma to anchor the glycocalyx.

• The glycocalyx is a layer of carbohydrates whose net negative charge prevents microbial attack and

mechanical damage caused by adhesion to neighboring RBCs or the endothelium.

• Glycolipids may bear copies of carbohydrate-based blood group antigens, such as antigens of the

ABH and the Lewis blood group systems.

RBC Membrane Proteins

• Although cholesterol and phospholipids constitute the principal RBC membrane structure,

transmembrane (integral) and cytoskeletal (skeletal, peripheral) proteins make up 52% of the

membrane structure by mass.

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• A proteomic study revealed there are at least 300 RBC membrane proteins, including 105

transmembrane proteins.

• Some proteins have a few hundred copies per cell, and others have more than a million copies per cell.

Of the purported 300 membrane proteins, about 50 have been characterized and named.

Transmembrane Proteins

• The transmembrane proteins serve many functions including transport sites, adhesion sites, and

signaling receptors.

• Any disruption in transport protein function changes the osmotic tension of the cytoplasm, which leads

to a rise in viscosity and loss of deformability.

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• Any change affecting adhesion proteins permits RBCs to adhere to one another and the vessel walls,

promoting fragmentation (vesiculation), reducing membrane flexibility, and shortening the RBC life

span.

• Signaling receptors bind plasma ligands and trigger the activation of intracellular signaling proteins,

which then initiate various energy-dependent cellular activities, a process called signal transduction.

• Through glycosylation, the transmembrane proteins also support surface carbohydrates, which join

with glycolipids to make up the protective glycocalyx.

• Most transmembrane proteins assemble into one of two major macromolecular complexes named by

their respective cytoskeletal anchorages: the ankyrin complex and the actin junctional complex, also

called protein 4.1 complex (Figure 6.3).

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• The anchoring of these transmembrane complexes to cytoskeletal proteins (adjacent to the inner or

cytoplasmic side of the membrane) prevents loss of the lipid bilayer.

• In addition, the linking of cytoskeletal proteins by the actin junctional complex provides membrane

structural integrity because the cell relies on an intact cytoskeleton to maintain its biconcave shape

despite deformability.

• Transmembrane proteins also provide vertical membrane structure.

• Blood group antigens. The blood group antigens are located in membrane macromolecular complexes

that serve as transporters, structural components, enzymes, receptors, and adhesion molecules.

• The carbohydrate-defined blood group antigens are supported in the RBC membrane by

transmembrane proteins.

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• Half of the known transmembrane proteins (approximately 25) are involved in the macromolecular

complexes that define blood antigen groups.

• More than 30 proteins bind to the GPI anchor and appear to float on the surface of the membrane.

Cytoskeletal Proteins

• The principal cytoskeletal proteins are the filamentous a-spectrin and b-spectrin which assemble to

form an antiparallel heterodimer held together with a series of lateral bonds.

• Antiparallel means that the carboxyl (COOH) end of one strand associates with the amino (NH3 ) end

of the other, and the two heterodimers self-associate head-to-head to form a tetramer.

• The ends of the spectrin tetramers are linked in the actin junctional complex, forming a hexagonal

cytoskeletal lattice adjacent to the inner (cytoplasmic) lipid bilayer.

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• This provides lateral or horizontal membrane stability.

• Because the cytoskeletal proteins do not penetrate the bilayer, they are also called peripheral

proteins.

• Dematin appears to stabilize the actin junctional complex and helps maintain the RBC shape.

• Membrane deformation. Spectrin dimer bonds that appear along the length of the molecules

disassociate and reassociate (open and close) during RBC deformation.

• These horizontal interaction defects inhibit the membrane’s ability to rebound from deformation.

• Ultimately, the RBCs progressively elongate to form visible elliptocytes, which causes mild to severe

hemolytic anemia.

• In a reduced surface area-to-volume ratio causes the formation of spherocytes.

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Osmotic Balance and Permeability

• The RBC membrane is impermeable to cations Na+, K+ , and Ca2+ .

• It is permeable to water and the anions bicarbonate (HCO3) and chloride (Cl2 ), which freely exchange

between plasma and RBC cytoplasm.

• Aquaporin 1 is a transmembrane protein that forms pores or channels whose surface charges create

inward water flow in response to internal osmotic changes.

• The ATP-dependent cation pumps Na+-ATPase and K+-ATPase regulate the concentrations of Na+

and K+ , maintaining intracellular-to-extracellular ratios of 1:12 and 25:1, respectively.

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• Ca2+-ATPase expels calcium from the cell, maintaining low intracellular levels of 30 to 60 nM

compared with 1.8 mM in the plasma.

• These pumps, in addition to aquaporin, maintain osmotic balance in the RBC.

• The cation pumps consume a significant portion of RBC ATP production.

• ATP loss or pump damage permits Ca2+ and Na+ influx, with water following osmotically. The cell

swells, becomes spheroid, and eventually ruptures. This phenomenon is called colloid osmotic

hemolysis.

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