Red Blood Cells Mature Erythrocytes Are Devoid of Internal
Organelles
- Anemia, a deficiency in the level of circulating hemoglobin (<
120-130 g/L)
Causes:
- genetic abnormalities (eg, sickle cell trait, pernicious
anemia), excessive bleeding, insufficiencies of dietary
iron or vitamin B 12, lysis of red blood cells by invading
pathogens (eg, malaria)
-Thrombocytopenia
Causes: bacterial infection, sulfa-containing antibiotics,
autoimmune reactions such as idiopathic
thrombocytopenic purpura, von Willebrand disease and
Glanzmann thrombasthenia
Red Blood Cells Derive From Hematopoietic Stem
Cells
 replacements are constantly being produced from
precursor stem cells
 self-renewal: possess a unique capacity both to
produce unaltered daughter cells
 Potency: to generate a diverse range of specialized
cell types
 stem cells are considered to exist in an
undifferentiated state
 Totipotent: capable of producing all the cells in an
organism
 pluripotent able to differentiate into cells of any of
the three germ layers
 multipotent able to produce only cells of a closely
related family
 Unipotent: able to produce only one type of cell.
 Stem cells are also classified as embryonic or adult
 Cytokines: glycoproteins that regulates
Differentiation of hematopoietic stem cells
 Stem cell factor, colony-stimulating factors
collaborate with interleukins 1, 3, and 6 to
stimulate the proliferation of hematopoietic stem
cells in the bone marrow
 Erythropoietin or thrombopoietin: directs
myeloid progenitor cells to eventually differentiate
into erythrocytes or platelets, respectively
 30-34 g/dL for an adult: amount of hemoglobin,
roughly one-third by weight in the interior of a red
blood cell.
mature enucleated red blood cells are unable to
reproduce.
 Red blood cells possess an extensive cytoskeletal
network responsible for maintaining their biconcave
configuration
1. disc-like configuration possesses a much higher
ratio of surface area to volume than more spherical
shapes.
2. enables red blood cells to fold over and squeeze
through capillaries
Erythrocytes Generate ATP Exclusively via Glycolysis
 Mature red blood cells lack the mitochondria that
contain ATP synthase and the enzymes of the
tricarboxylic acid cycle, electron transport chain,
and β-oxidation pathway.
 incapable of utilizing fatty acids or ketone bodies as
metabolic fuel
 completely reliant of glycolysis to generate ATP.
 facilitated diffusion: Glucose enters red blood cells
in this process
 mediated by glucose transporter 1 (GLUT1), also
known as glucose permease
 glycolytic pathway in red blood cells also possesses
a unique branch, or shunt, whose purpose is to
isomerize 1,3-bisphosphoglycerate (1,3-BPG) to
2,3-bisphosphoglycerate (2,3-BPG).
 2,3-BPG binds to and stabilizes hemoglobin in the
T-state
 2,3-BPG mutase: catalyze Conversion of 1,3-BPG
to 2,3-BPG
 also catalyzes the hydrolysis of 2,3-BPG to the
glycolytic intermediate 3 phosphoglycerate
 multiple inositol polyphosphate phosphatase: A
second enzyme,catalyzes the hydrolysis of 2,3-BPG
to the glycolytic intermediate 2
bisphosphoglycerate.
 Erythrocytes: high levels of the enzyme carbonic
anhydrase. enables them both to absorb waste CO2
by catalyzing its rapid conversion to carbonic acid,
 reverse this process in order to facilitate
its expulsion in the lungs
 red blood cells carry some CO2 in the form of
hemoglobin-bound carbamates
 80% is carried as dissolved carbonic acid and
bicarbonate.
RED BLOOD CELLS MUST BE CONTINUALLY
REPLACED
 About Two Million Red Blood Cells Enter the
Circulation per Second
 120-day lifespan of a normal red blood cell requires
that nearly 1% of the 20 to 30 trillion erythrocytes
individual must be replaced daily.
 nascent red blood cells, called reticulocytes, retain
the capacity to synthesize polypeptides (eg, globin)
under the direction of vestigial mRNA molecules
 Diamond-Blackfan anemia: mutations in the gene
encoding for the ribosomal processing protein
RPS19.
 5q-syndrome: is caused by mutations that lead to an
insufficiency of ribosomal protein RPS14.
Erythropoietin Regulates Production of Red Blood
Cells
 Erythropoiesis: The initial stages are the
production of red blood cells, are modulated by
1. stem cell factor
2. colonystimulating factors
3. interleukins 1, 3, and 6.
 myeloid progenitor cells to differentiation into
erythrocytes is largely dependent on erythropoietin
(EPO)
 a glycoprotein of 166 amino acids
 EPO: synthesized mainly by the kidney, is
released into the bloodstream in response to
hypoxia.
 Upon reaching the bone marrow, EPO stimulates red
blood cell progenitors via a trans membrane
receptor.
 Binding of EPO to its receptor stimulates the
dimerization of the receptor and activation of
associated molecules of the Jak2 protein-tyrosine
kinase.
 Methemoglobin: Hemoglobin in which one or more
heme irons has been oxidized to the ferric (Fe3+)
state
 Ferric hemes do not bind oxygen: reduces the
number of O 2 binding sites,
 methemoglobin reduction is catalyzed by the
NADH–cytochrome b 5 methemoglobin reductase
system. The first component of the system,
 cytochrome b 5reductase: a flavoprotein named
(also known as methemoglobin reductase),
transfers electrons from NADH to the second
component, cytochrome b5, using electrons supplied
by NADH:
 ultimate source of the electrons used to reduce
methemoglobin is glycolysis
 NAD + is reduced to NADH by the action of
glyceraldehyde-3-phosphate dehydrogenase
 Methemoglobinemia: the abnormal accumulation
of methemoglobin, can arise from genetic
abnormalities (inherited methemoglobinemia) or
(acquired methemoglobinemia) ingestion of
chemicals such as sulfonamides or aniline
 Bluish discoloration/ cyanosis
 membrane-specific : Mutations that affect the
cytoskeletal proteins responsible for maintaining
their biconcave shape and resistance to osmotic
pressure
 Ex. hereditary spherocytosis and hereditary
elliptocytosis,

 Inhirited form: diffeciency of cytochrome B5  arise from abnormalities in the amount or structure
reductase. of the cytoskeletal protein spectrin

 moglobinemia can result from mutations in
hemoglobin itself,
 affecting the proximal and distal histidine
residues
Superoxide Dismutase, Catalase, & Glutathione:
Protect Blood Cells From Oxidative Stress & Damage
Paroxysmal Nocturnal Hemoglobinuria: Defects in the
synthesis of the glycophosphatidylinositol groups that
anchor acetylcholinesterase and decay-accelerating
factor, to the surface of the erythrocyte membrane.
THE RED BLOOD CELL MEMBRANE
 10 major proteins

 the polypeptide with the highest molecular mass,

 3% of the hemoglobin of human blood undergoes
auto-oxidation each day.
 NADPH-hemoprotein reductase (a cytochrome
P450 reductase,another enzyme that can catalyze the
reduction of the Fe3+ in methemoglobin to Fe2+.
 red blood cells renders them completely reliant on
the pentose phosphate pathway
 the X-linked enzyme glucose-6-phosphate
dehydrogenase for the reduction of NADP+ to
NADPH.
which migrates slowest, was designated band 1
protein, also known as spectrin
 The Red Blood Cell Membrane Contains Anion
Exchange Protein & the Glycophorins
 Band 3 protein is a multipass transmembrane
glycoprotein whose polypeptide chain crosses the
bilayer 14 times.
 oriented with its carboxyl terminal end: from
external surface
 amino-terminal end: cytosolic face.

 Glucose-6-phosphate dehydrogenase deficiency is

the most common of all Enzymopathies

 400 million people are estimated to carry one of the

over 140 genetic variants of glucose-6-phosphate
dehydrogenase

 glucose-6-phosphate dehydrogenase deficiency:

renders red blood cells hypersensitive to oxidative
stress and ROS-induced the formation of Heinz
bodies, insoluble aggregates consisting of
hemoglobin molecules whose —SH groups have
become oxidized

 Extrinsic causes (beyond the erythrocyte membrane)

include hypersplenism : enlargement of the spleen
causes red blood cells to become sequestered within
this organ

 Escherichia coli and clostridia, secrete lytic factors

called hemolysins

 The root cause of many hemolytic anemias is

intracellular, also referred to as intrinsic.

Ex: Gluocse-6-phosphate dehydrogenase deficiency

 principal function of this dimeric anion exchange
protein: provide a channel through the membrane
via which chloride and bicarbonate anions can be
exchanged
 bicarbonate generated from the hydration of CO2 is
exchanged for chloride
 Glycophorins A, B, and C are single-pass
transmembrane proteins (the polypeptide chain
crosses the membrane only once
 glycophorin A, is comprised of a 131-amino acid
polypeptide covalently modified by 16
oligosaccharide chains,
 15 of them O-linked, that account for roughly 60%
of its mass and nearly 90% of the sialic acid residues
exposed on the surface of the red cell membrane
 carboxyl-terminal end extends into the
cytosol and binds to band 4.1 protein,
which in turn binds to spectrin.
 Polymorphism of glycophorin A serves as the basis
of the MN blood group system
 influenza virus and Plasmodium falciparum, target
erythrocytes by recognizing and binding to
glycophorin A.
 individuals whose red cells lack glycophorin A
exhibit no adverse effects.
Spectrin, Ankyrin, & Other Peripheral Membrane Proteins
Help Determinethe Shape & Flexibility of the Red Blood
Cell
 Has flexible network of cytoskeletal proteins
 Spectrin is the most abundant protein of the
erythrocyte cytoskeleton.
 It is composed of two polypeptides more than
2100 residues in length
 spectrin 1 (α chain) and spectrin 2 (β chain).
 The α and β chains of each spectrin dimer
intertwine in an antiparallel orientation to form
a highly extended structural unit
 two spectrin dimers self-associate
 head-to-head linked to the inner surface of the
plasma membrane (and is bridged to other
spectrin tetramers) via ankyrin
 Ankyrin is a pyramid-shaped protein that binds
spectrin.
 binds tightly to band 3, securing attachment of
spectrin to the membrane.
 sensitive to proteolysis
 Actin (band 5) exists in red blood cells as short,
doublehelical filaments of F-actin.
 - tail end of spectrin dimers binds to actin.
 -Actin also binds to protein 4.1.
 -Protein 4.1 is a globular protein that binds tightly
to a site near the actinbinding domain in the tail of
spectrin to form a protein 4.1-spectrin-actin
ternary complex. Protein 4.1 also binds to the
integral membrane proteins glycophorin A and
glycophorin C, as well as certain phospholipids,
anchoring the
 ternary complex to the membrane.
Abnormalities in the Amount or Structure of
Spectrin Cause Hereditary Spherocytosis &
Elliptocytosis
 Hereditary spherocytosis: genetic disease
transmitted as autosomal dominant
 characterized by the presence of spherocytes
(spherical red blood cells) by
 hemolytic anemia, and by splenomegaly. renders
spherocytes less deformable and more prone to
destruction in the spleen
 Short life span in circulation.
 -affects about 1 in 5000 persons of Northern
European ancestry
 -caused by a deficiency in the amount or
abnormalities in the structure of spectrin or less
frequently, ankyrin or band 3, 4.1, or 4.2 proteins.
 The anemia associated with hereditary spherocytosis
is generally relieved by surgical removal of the
patient’s spleen (splenectomy)
 Hereditary elliptocytosis: red blood cells assume
an elliptic shape. genetic abnormalities that affect
spectrin or, less frequently, band 4.1 protein or
glycophorin C.
 The term “blood type” refers to the antigenic
phenotype, usually recognized by the use of
appropriate antibodies
 -type A have anti-B antibodies in their plasma that
will agglutinate the erythrocytes in type B or type
AB blood.
 - type B have anti-A antibodies that will agglutinate
type A or type AB erythrocytes. -Type AB blood
 has neither anti-A nor anti-B antibodies, and has
been designated the universal recipient.
 -Type O blood has neither A nor B antigens, and
has been designated the universal donor.
 -The genes responsible for production of the ABO
substances are present on the long arm of
chromosome 9.
 There are three alleles, two of which are
codominant (A and B) and the third (O) recessive;
 determine which of the four phenotypic products is
synthesized: the A, B, AB, and O substances.
The ABO Antigens Are Glycosphingolipids &
Glycoproteins
 ABO antigens are complex oligosaccharides present
in most cells of the body and in certain secretions
 bound to membrane proteins or lipids, and are
collectively referred to as ABO substances.
 RBC: the membrane oligosaccharides that
determine the antigenic natures of the ABO
substances appear to be mostly present in
glycosphingolipids, whereas in secretions
 the same oligosaccharides are present in
glycoproteins.
The A Gene Encodes a GalNAc Transferase, the B Gene a Gal
Transferase, & the O Gene an Inactive Product
 H substance, the blood group substance found in
persons of type O, is the precursor of both the A and
B substances formed by the action of a
fucosyltransferase,
 coded for by the H locus, that catalyzes the addition
of an α1 → 2
 A substance contains an additional GalNAc, while
B substance contains an additional Gal
 The A gene encodes a UDP-GalNAc-specific
GalNAc transferase that adds the terminal GalNAc
to H substance.
 The B gene encodes a UDP-Gal-specific Gal
transferase that adds the Gal residue to H
substance.
 type AB possess both enzymes, and thus synthesize
two oligosaccharide chains
 all individuals carrying the hh genotype will have
red blood cells of type O, referred to as the Bombay
phenotype (Oh)
Platelets Contain Mitochondria,
But Lack a Nucleus
 -When megakaryocytes, the progenitors of red blood
cells, are exposed tothrombopoietin they may
fragment and form platelets
 possess mitochondria, lysozymes, and a tubular
network that forms an open canalicular system.
 -spheroidal at rest, thereby facilitating the secretion
of various endocrine and coagulation factors
 factors are stored inside the platelets within densely
packed secretory vesicles, called dense granules,
(contain Ca2+, ADP, and serotonin
 α-granules: contain fibrinogen, fibronectin,
platelet-derived growth factor, von Willebrand
factor, or other coagulation factors that are released
on receipt of an appropriate stimulus
 circulate at a density of 2 to 4 × 105 platelets per
milliliter of blood.
 Platelets: derive the majority of their energy from
metabolizing glucose,
 their mitochondria enable them to generate ATP via
the β-oxidation of fatty acids.
 Immune thrombocytopenic purpura is an
autoimmune disorder marked by depressed platelet
counts (thrombocytopenia) caused by the
generation of antibodies against the patient’s own
platelets
 Thrombocytopenia also can occur when persons
who are homozygous for a mutant variant of
glycoprotein IIb/IIIa in which the leucine 33 is
replaced by proline receive blood from a donor that
is homo- or heterozygous for the wild-type form of
this major platelet antigen
 neonatal alloimmune thrombocytopenia, which
affects roughly 1 in 200 term pregnancies
 -antibodies from the mother’s circulation cross the
placental barrier and attack platelets in the fetus’
circulatory system
 can be induced by drugs such as tamoxifen,
ibuprofen, vancomycin, and sulfonamides.
 hemolytic-uremic syndrome: a disease of infants
characterized by progressive kidney failure, include
both thrombocytopenia and hemolytic anemia.
 von Willebrand disease: the abnormal bleeding
associated with a genetic defect that compromises
the ability of platelets to adhere to the endothelium,
rather than a deficit in platelet number.
 Bernard-Soulier syndrome: genetically inherited
deficiency in glycoprotein 1b

 Glanzmann thrombasthenia: genetically inherited

deficiency in the glycoprotein IIb/IIIa complex).

 The first pathophysiologic condition to be treated by

gene therapy was a deficiency of adenosine
deaminase.

Summary
 2,3-Bisphosphoglycerate mutase catalyzes the 
isomerization of the glycolytic intermediate 1,3-
bisphosphoglycerate to form the 2,3-
bisphosphoglycerate, which stabilizes T-state

 Cytochrome b5 reductase reduces the Fe3+ of

methemoglobin to Fe2+, restoring function.

 The red cell contains a battery of cytosolic enzymes

— superoxide dismutase, catalase, and
glutathione peroxidase— that catalyze the
neutralization of reactive oxygen species.

 Deficiencies in the quantity or the activity of

glucose-6- phosphate dehydrogenase, which
produces NADPH, constitute a major cause of
hemolytic anemia.

 Cytoskeletal proteins such as spectrin, ankyrin, and

actin interacting with integral membrane proteins
underlie the flexible biconcave shape of red blood
cells.

 Band 4.1 protein facilitates the exchange of

bicarbonate and chloride ions across erythrocyte
membranes.

 The ABO blood group substances of the red cell

membrane

 are complex glycosphingolipids. The

immunodominant sugar

 of A substance is N-acetylgalactosamine, whereas

that of B

 substance is galactose. O substance contains

neither of these

 sugar residues.

 ■ Plateletsare small, enucleated fragments of

larger precursor megakaryocytes