ERYTHROCYTE METABOLISM

AND
MEMBRANE STRUCTURE
RED BLOOD CELL
• the primary blood cell circulating at 5 million cells/microliter of blood on
average
• anucleate, biconcave, and has an average volume of 90 fL
• contains hemoglobin that consists of four globin molecules, each
supporting one heme molecule and each heme molecule contains a
molecule of iron
• Cytoplasmic ribosome and mitochondria disappear 24 - 48 hours after
bone marrow release, eliminating the cell’s ability to produce proteins or
support oxidative metabolism.
• Energy (ATP) is produced within the cytoplasm through anaerobic
glycolysis for the lifetime of the cell.
• Glycolysis
• Process that converts glucose to pyruvate for energy purposes
• Gluconeogenesis
• Formation of glucose from non-carbohydrate sources (amino acids,
lactate, glycerol [lipids])
• Glycogenolysis
• Breakdown of glycogen to produce glucose
• Glycogenesis
• Conversion of glucose to glycogen for storage purposes
ENERGY PRODUCTION–ANAEROBIC
GLYCOLYSIS
• Anaerobic glycolysis (Embden-Mayerhof Pathway [EMP]) requires
glucose to generate ATP.
• Glucose (from plasma) enters the RBC without energy expenditure
via the transmembrane protein Glut-1.
• Through the EMP, glucose is catabolized to pyruvate, consuming 2
molecules of ATP per molecule of glucose and maximally
generating 4 molecules of ATP per molecule of glucose, for a net
gain of 2 molecules of ATP.
EMBDEN-MEYERHOF PATHWAY
EMBDEN-MEYERHOF PATHWAY
• Major glycolytic pathway in the RBC
• Handles 90% of glucose utilization in the RBC
• Non-oxidative, anaerobic pathway
• Produces a net gain of 2 ATP molecules
• ATP in the RBC is used in the following ways:
• Maintenance of RBC shape and deformability
• Gives energy for the active transport of cations
• Helps in modulating the amount of 2,3 bisphosphoglycerate (2,3-BPG) generated
• Pyruvate kinase (PK) deficiency – most common enzyme deficiency of
EMP and is the most common form of HNSHA
GLYCOLYSIS DIVERSION PATHWAYS
(SHUNTS)
• Three alternative pathways, called diversions or shunts, branch
from the glycolytic pathway:

• Hexose Monophosphate Pathway (HMP) or aerobic glycolysis

• Methemoglobin Reductase Pathway

• Rapoport-Leubering Pathway
HEXOSE MONOPHOSPHATE PATHWAY
• Aerobic or oxidative glycolysis
• Also known as pentose phosphate
shunt and phosphogluconate pathway
• Detoxifies peroxide which arise from
oxygen reduction in the cells aqueous
environment, where it oxidizes and
destroys heme iron, proteins, and
lipids, especially lipids containing
thiol groups.
• By detoxifying peroxide, HMP
extends the functional life span of
the RBC
HEXOSE MONOPHOSPHATE PATHWAY
• During steady-state glycolysis, 5%-10% of G6P is diverted to the
HMP.
• The activity of HMP may rise up to thirtyfold during oxidative
challenge.
• Prevents the denaturation of globin by oxidation
• Only G6PD provides the means of generating NADPH for
glutathione reduction.
• G6PD deficiency, is the most common RBC enzyme deficiency
worldwide, results in HNSHA
METHEMOGLOBIN REDUCTASE PATHWAY
• Peroxide oxidizes heme iron from the ferrous (+2) to the ferric (+3)
state (methemoglobin).
• Although HMP prevents hemoglobin oxidation by reducing
peroxide, it is not able to reduce methemoglobin once it’s formed.
• NADPH is able to slowly reduce methemoglobin. This activity is
enhanced by methemoglobin reductase
• Also called cytochrome b5 reductase, that accounts for more than
65% of the methemoglobin-reducing capacity inside the RBC.
RAPOPORT-LEUBERING PATHWAY
• Generates 2,3-bisphosphoglycerate (2,3-BPG)
• 2,3-BPG regulates oxygen delivery to the tissues by competing
with oxygen from the oxygen-binding site of hemoglobin.
• This diversion sacrifices the production of 2 ATP molecules.
• Acidic pH and low concentrations of 3-PG and 2-PG inhibit the
activity of bisphosphoglycerate mutase.
• The above mentioned conditions plus decreased ATP levels
activate bisphosphoglycerate phosphatase.
RBC MEMBRANE
• RBC MEMBRANE DEFORMABILITY
• RBCs are biconcave, average 90 fL in volume, average 140 m surface area
(40% more than 90 fL sphere), able to stretch undamaged up to 2.5 times
its diameter as it pass through splenic pores 2 m in diameter.
• RBC plasma membrane (5 m thick) is 100 times more elastic than
comparable latex membrane but has tensile strength than that of steel.
• RBC deformability not only depends on RBC geometry but also on relative
cytoplasmic (hemoglobin) viscosity.
RBC MEMBRANE
• RBC MEMBRANE LIPIDS
• Elasticity (pliancy) also contribute to deformability
• The RBC membrane consists of approximately 8% carbohydrates, 52%
proteins, and 40% lipids.
• The lipid portion that consists of equal parts of cholesterol and
phospholipids form a bilayer.
• Phospholipids form an impermeable fluid barrier as their hydrophilic polar
head groups are arrayed upon the membrane’s surface, oriented toward
both the aqueous plasma and cytoplasm, respectively and their
hydrophobic nonpolar acyl tails arrange themselves to form a central layer
dynamically sequestered from the aqueous plasma and cytoplasm.
RBC MEMBRANE
• RBC MEMBRANE LIPIDS
• Cholesterol confers tensile strength to the lipid bilayer.
• The ration of cholesterol to phospholipid remains relatively constant and
balances the need for deformability and strength.
• Phospholipids are asymmetrically distributed:
• Phosphatidylcholine and sphingomyelin predominate the outer later
• Phosphatidylserine (PS) and phosphatidylethanolamine form most of the inner layer
• Distribution of these phospholipids is energy dependent, relying on a number of
membrane-associated enzymes (flippases, floppases, and scramblases)
• When this distribution is disrupted, PS (the only negatively charged phospholipid)
redistributes to the outer layer. Splenic macrophages possess receptors that bind
PS and destroy senescent and damaged RBCs.
• Glycocalyx – a layer of carbohydrates whose net charge prevents microbial attack
and protects the RBC from mechanical damage caused by adhesion to
neighboring RBCs or to the endothelium.
RBC MEMBRANE
• RBC MEMBRANE PROTEINS
• Transmembrane proteins (integral proteins)
• Channel ions, water, and glucose
• Anchor cell membrane receptors
• Provide the vertical support connecting the lipid bilayer to the underlying
cytoskeleton to maintain membrane integrity
• Through glycosylation transmembrane proteins support surface
carbohydrates, which join with glycolipids to make up the protective
glycocalyx
• Signal transduction – process of binding of signaling receptors to plasma
ligands that trigger activation of intracellular signaling proteins which
initiate various energy-dependent cellular activities
RBC MEMBRANE
• RBC MEMBRANE PROTEINS
• Cytoskeletal proteins (peripheral proteins)
• Do not penetrate the bilayer
• Provide the horizontal or the lateral support for the membrane
• The shape and flexibility of the erythrocyte depends on the cytoskeleton
RBC MEMBRANE
• OSMOTIC BALANCE AND PERMEABILITY
• RBC membrane is impermeable to cation Na+, K+, and Ca2+ but permeable to
water and the anions HCO3- and Cl-, which freely exchange between the plasma
and RBC cytoplasm.
• Aquaporin 1 forms pores or channels whose surface charges create inward water
flow in response to internal osmotic changes.
• Na+-ATPase and K+-ATPase are ATP-dependent cation pumps that regulate the
concentrations of Na+ and K+, maintaining intracellular-to-extracellular ratios of
1:12 and 25:1, respectively.
• Ca+2-ATPase extrudes calcium, maintaining low intracellular levels to 5-10 mol/L.
• Calmodulin, a cytoplasmic Ca2+-binding protein, controls the function of Ca2+-
ATPase
END