BLOK BIOMEDIK II FK UHO KENDARI

SEMESTER GANJIL TA. 2018/ 2019

Ika Yustisia
Departemen Biokimia
Fakultas Kedokteran UNHAS
 Definisi, fungsi, dan komposisi darah
 Protein plasma
 Eritropoietin, trombopoietin, dan sitokin
yang berperan pada hematopoiesis
 Struktur dan fungsi spesifik membran
eritrosit
 Dasar biokimiawi penggolongan darah ABO
 Hemoglobin dan metabolisme besi
 Metabolisme spesifik eritrosit dan trombosit
 Hemostasis
 Red fluid that circulates in the
heart, arteries, capillaries, and
veins carrying nourishment and
oxygen to and bringing away
waste products from all parts of
the body
As the primary avenue by which tissues
are connected to each other and the
surrounding environment, the blood
that circulates throughout our body
performs a variety of functions.
 Human blood constitutes
about 8% of the
body’sweight
 Consists of cells and cell
fragments in an aqueous
medium (= the blood
plasma)
 The proportion of cellular
elements (=hematocrit)
in the total volume is
approximately 45%
 Plasma Serum

Anti-coagulated blood Coagulated blood

No need anti-
Need anti-coagulants
coagulants

Fibrinogen is present Fibrinogen is absent

Plasma = serum + fibrinogen and other

clotting factors
 The blood plasma is an aqueous solution of
electrolytes, nutrients, metabolites,
proteins, vitamins, trace elements, and
signaling substances
 The fluid phase of coagulated blood is
known as blood serum
 Serum differs from the plasma in that it
lacks fibrin and other coagulation proteins
 Quantitatively,
proteins are the most
important part of the
soluble components of
the blood plasma
 With concentrations of
between 6 and 8 g/dL,
they constitute
approximately 4% of
the body’s total
protein
 Some 100 different proteins occur in
human blood plasma.
 Based on their behavior during
electrophoresis they are broadly divided
into five fractions: albumins and α1–, α2–,
β– and γ-globulins
Albumin is the major
protein in human
plasma
 Roughly 70% to 80% of
all plasma proteins are
synthesized in the liver.
 These include albumin,
fibrinogen, transferrin,
and most components of
the complement and
blood coagulation
cascade
 One prominent exception
is the γ-globulins, which
are synthesized in the
lymphocytes
 The liver synthesizes
approximately 12 g of albumin per
day
 About 25% of total hepatic protein
synthesis and half its secreted
protein.
 About three-fifths of total plasma
protein by weight (3.4-4.7 g/dL)
 The remainder resides in the
extracellular space.
 Responsible for 75% to 80% of the
osmotic pressure of human plasma
 Albumin is also a major transporter
 The various types of cells (lineages) that make up the
blood are constantly being produced in the bone
marrow.
 All cell lineages are descended from hematopoietic stem
cells, cells that are renewable throughout the life of the
host.
 The population of hematopoietic stem cells is quite small.
Estimates vary between 1 and 10 per 105 bone marrow
cells.
 In the presence of the appropriate signals, hematopoietic
stem cells proliferate, differentiate, and mature into any
of the types of cells that make up the blood
 Cytokines are small secreted proteins released
by cells have a specific effect on the interactions
and communications between cells.
 Developing progenitor cells in the marrow grow
in proximity with marrow stromal cells
(fibroblasts, endothelial cells, adipocytes, and
macrophages)
 Thestromal cells form an extracellular matrix
and secrete growth factors (=cytokines) that
regulate hematopoietic development.
The 120 day lifespan
of a normal red
blood cell requires
that nearly 1% of
the roughly 30 trillion
erythrocytes in a typical
individual must be replaced daily.
This equates to a rate of
production of ∼2 million new red
blood cells per second.

The production of red cells is regulated by the demands of oxygen delivery to the
tissues. In response to reduced tissue oxygenation, the kidney releases the hormone
erythropoietin, which stimulates the multiplication and maturation of erythroid
progenitors. Erythropoietin is a glycoprotein of 166 amino acids (34 kDa)
  A potent cytokine that
regulates
megakaryocyte and
platelet production.
 Produced primarily in
the liver but also in the
bone marrow and
kidney
 Binds to the TPO
receptor (TPO-R)
 Acts at all stages of
thrombopoiesis to
regulate the
development and
maturation of
megakaryocytes and
subsequent release of
Thrombopoietin platelets
 Under the microscope, the red blood cell appears
to be a red disc with a pale central area
(biconcave disc)
 The biconcave disc shape serves to facilitate gas
exchange across the cell membrane
 The membrane proteins:
• Maintain the shape of
the red blood cell
• Allow the red blood cell
to traverse the capillaries
with very small luminal diameters
to deliver oxygen to the tissues
 The interior diameters of many capillaries
are smaller than the approximately 7.5μm
diameter of the red cell
 The spleen (=limpa) is the organ
responsible for determining the viability
of the red blood cells
 Erythrocytes pass through the spleen 120
times per day
 The elliptical passageways through the
spleen are approximately 3μm in diameter,
and normal red cells traverse them in
approximately 30 seconds
 Damaged red cells that are no longer
deformable become trapped in the
passages in the spleen, where they are
destroyed by macrophages
 On the cytoplasmic side of the
membrane, proteins form a two-
dimensional lattice that gives the red cell
its flexibility
 The major proteins are spectrin, actin,
band 4.1, band 4.2, and ankyrin.
 When the red blood cell is subjected to mechanical stress, the
spectrin network rearranges. Some spectrin molecules
become uncoiled and extended; others become compressed,
thereby changing the shape of the cell, but not its surface
area.
 The major protein,
spectrin, is linked to the
plasma membrane either
through interactions with
ankyrin and band 3, or with
actin, band 4.1, and
glycophorin
 The cytoskeleton of
erythrocyte displays the
cross-linking of the
spectrin dimers to actin
and band 3 anchor sites.
 Approximately 30 human blood group
systems have been recognized, the best
known of which are the ABO, Rh (Rhesus),
and MN systems.
 The term “blood type” refers to the
antigenic phenotype, usually recognized
by the use of appropriate antibodies.
 The ABO system
 Discovered by Karl Landsteiner (1900)
 The membranes of the erythrocytes of
most individuals contain one blood group
substance of type A, type B, type AB, or
type O.
Agglutinogen

Agglutinin
 The human ABO blood groups illustrate
the effects of glycosyltransferases on
the formation of glycoproteins.
 Each blood group is designated by the
presence of one of the three different
carbohydrates, termed A, B, or O,
attached to glycoproteins and
glycolipids on the surfaces of red blood
cells.
 The ABO blood group antigens are
encoded by one genetic locus, the ABO
locus
 The ABO locus encodes specific
glycosyltransferases that synthesize A
and B antigens on RBCs.
 The ABO locus is located on chromosome 9
at 9q34.1-q34.2.
 It contains 7 exons that span more than 18 kb
of genomic DNA.
 Exon 7 is the largest and contains most of the
coding sequence.
 Exon 6 contains the deletion (deletion of
guanine at position 261) that is found in most
O alleles and results in a loss of enzymatic
activity.
 The A and B alleles differ from each
other by seven nucleotide substitutions,
four of which translate into different
amino acids in the gene product:
R176G, G235S, L266M, G268A
 The residues at positions 266 and 268
determine the A or B specificity of the
glycosyltransferase they encode.
 The most important task of erythrocytes
is to transport molecular oxygen (O2)
from the lungs into the tissues, and
carbon dioxide (CO2) from the tissues
back into the lungs
 To achieve this, the higher organisms
require a special transport system, since
O2 is poorly soluble in water
 Only around 3.2 mL O2 is soluble in 1 L
blood plasma.
 By contrast, the protein hemoglobin (Hb),
contained in the erythrocytes, can bind a
maximum of 220 mL O2 per liter—70
times the physically soluble amount
 The Hb content of blood:
• 14 – 18 g/dL in men
• 12 – 16 g/dL in women
 Adult hemoglobin is a tetrameric
hemeprotein (quartenery structure) found
in erythrocytes where it is responsible for
binding oxygen in the lung and
transporting the bound oxygen
throughout the body where it is used in
aerobic metabolic pathways
 The tetramers are composed of pairs of
two different polypeptide subunits
 The subunits are β-, γ-, α-, δ-chain
The haemoglobin molecule is a tetramer consisting of 4 polypeptide
chains, known as globins, which are usually:
• 2 alpha chains that are each 141 amino acids long
• 2 beta chains that are each 146 amino acids long
Attached to each chain is an iron-containing molecule known as heme
 The forms of adult hemoglobin are
• HbA1: α2β2 (97%)
• HbA2: α2δ2 (2-3%)
 Two other forms occur during embryonic and
fetal development
• The first three months, embryonic hemoglobins are
formed, with the ζ2ε2 and α2ε2
• Up to the time of birth, fetal hemoglobin then
predominates (HbF, α2γ2)
• During the first few months of life, it is replaced gradually
by HbA
 After red blood cells reach the end of their
life span (approximately 120 days), they are
phagocytosed by cells of the
reticuloendothelial system.
 Globin is cleaved to its constituent amino
acids, and iron is returned to the body’s iron
stores.
 Heme is oxidized and cleaved to produce
carbon monoxide and biliverdin -----------
recall Porphyrin Metabolism (Blok Biomedik 1)
 Mature erythrocytes contain no
intracellular organelles, so the
metabolic enzymes of the red blood cell
are limited to those found in the cytoplasm
 The cytosol of the red blood cell contains
enzymes necessary for the prevention
and repair of damage done by reactive
oxygen species and the generation of
energy
 Erythrocytescan only generate adenosine
triphosphate (ATP) by glycolysis
 The ATP is used for:
• Ion transport across the cell membrane (primarily
Na+, K+, and Ca2+)
• The phosphorylation of membrane proteins
• The priming reactions of glycolysis

 Erythrocyte
glycolysis also uses the
Rapaport-Luebering shunt to generate 2,3-
bisphosphoglycerate (2,3-BPG)
Glycolysis is the major pathway,
with branches for the hexose
monophosphate shunt (for
protection against oxidizing agents/
ROS) and the Rapoport–Luebering
shunt (which generates 2,3-
bisphosphoglycerate, which
moderates oxygen binding to
hemoglobin).

The NADH generated from Due to their role in O2

glycolysis can be used to reduce transport, the erythrocytes
methemoglobin (Fe3+) to normal are constantly exposed to
hemoglobin (Fe2+), or to convert high concentrations of O2
pyruvate to lactate and are therefore
particularly at risk from ROS
 Thrombocytes are cytoplasmic fragments that are
released by megakaryocytes in the bone marrow
and stored in the spleen.
 Do not have nuclei, but contain a number of
organelles such as mitochondria, lysosomes and
peroxisomes.
 Play an important role in hemostasis and are essential
for thrombus formation at sites of injury.
 With a lifetime of 5–7 days in the circulation and no
nucleus their metabolic program must be stable over
this time period and be available for the energy
requiring processes engaged when they are activated.
 Both mitochondrial oxidative phosphorylation
and glycolysis are highly active in platelets
 It has been estimated that in the resting platelet,
65% of the ATP is generated from glycolysis and
35% from oxidative phosphorylation
 Mitochondrial fatty acid oxidation can
contribute to ATP production in platelets in both
the resting and thrombin stimulated state.
 Platelets contain three types of granules:
 Type I (electron-dense granule) contains calcium,
adenosine diphosphate (ADP), adenosine triphosphate
(ATP), and serotonin.
 Type II (α-granule) contains a heparin antagonist,
platelet-derived growth factor, β-thromboglobulin,
fibrinogen, von Willebrand factor (vWF), and other clotting
factors.
 Type III (lysosomal granule) contains hydrolytic
enzymes.
During activation, the contents of these granules, which
modulate platelet aggregation and clotting, are secreted
 Following injury to blood vessels,
hemostasis ensures that blood loss is
minimized.
 Three phases of hemostasis:
• Phase 1: Procoagulation
• Phase 2: Anticoagulation
• Phase 3: Fibrinolysis
 Phase 1 (procoagulation) leads to
production of fibrin from fibrinogen and
aggregation into an insoluble network
(=clot), which covers the rupture area and
prevents further loss of blood
 Platelets are responsible to form
mechanical plugs at the site of vessel
injury and to secrete regulators of the
clotting process and vascular repair
Three fundamental mechanisms are involved
in platelet function during blood
coagulation:
Mechanism Action
The platelet subendothelial
Adhesion interaction

Shape change, long pseudopods,

Aggregation platelet-platelets interaction

Secretion Release the contents of granules

Adhesion sets off a series of reactions termed platelet
activation, which leads to platelet aggregation and
secretion of platelet granule contents.
 Thrombus (clot) formation is enhanced
by thrombin activation, which is mediated
by the complex interaction that constitutes
the blood coagulation cascade.
 This cascade consists primarily of
proteins that serve as enzymes or
cofactors, which function to accelerate
thrombin formation and localize it at the
site of injury.
Cleavage of fibrinogen results in
clot formation.
A. Fibrinogen, the precursor
protein of fibrin, is formed
from two triple helices joined
together at their N-terminal
ends
B. Thrombin, a serine protease,
cleaves the terminal portions
of fibrinogen that contain
negative charges. The fibrin
monomers can then
aggregate and form a “soft”
clot.
Hard clots – cross-linking of fibrin
Factor XIIIa catalyzes a
transamidation reaction between
glutamine and lysine side chains
on adjacent fibrin monomers.
The covalent cross-linking takes
place in three dimensions,
creating a strong network of fibers
that is resistant to mechanical and
proteolytic damage.
This network of fibrin fibers traps
the aggregated platelets and other
cells, forming the clot that plugs
the vent in the vascular wall
Vitamin K-dependent formation of γ-carboxyglutamate residues. Thrombin, factor VII, factor IX,
and factor X are bound to their phospholipid activation sites on cell membranes by Ca2+. The
vitamin K-dependent carboxylase, which adds the extra carboxyl group, uses a reduced form of
vitamin K (KH2) as the electron donor and converts vitamin K to an epoxide.
 Toprevent the coagulation reaction
from becoming excessive, the blood
contains a number of anticoagulant
substances, including highly
effective proteinase inhibitors.
 Antithrombin III binds to various serine proteinases
in the cascade and thereby inactivates them.
 Heparin, an anticoagulant glycosaminoglycan
potentiates the effect of antithrombin III.
 The ATIII–heparin complex can inactivate thrombin
and the serine protease factors XIIIa, XIa, IXa, and Xa
 Thrombomodulin, which is located on the vascular
endothelia, also inactivates thrombin.
 A glycoprotein known as Protein C ensures
proteolytic degradation of factors V and VIII.
 The fibrin thrombus resulting from blood clotting is
dissolved again by plasmin, a serine proteinase found
in the blood plasma.
 For this purpose, the precursor plasminogen first has
to be proteolytically activated by enzymes from
various tissues.
 This group includes the plasminogen activator from
the kidney (urokinase) and tissue plasminogen
activator(t-PA) from vascular endothelia.
 By contrast, the plasma protein α2-antiplasmin, which
binds to active plasmin and thereby inactivates it,
inhibits fibrinolysis.
 Murray, RK., Granner, DK., Rodwell,
VW(editors). Harper’s Ilustrated
Biochemistry,), 30th ed., 2015
 Lieberman, M., Marks, DA. Marks’ Basic
Medical Biochemistry a Clinical Approach 4rd
ed. Lippincott Williams and Wilkins, 2013
 Meisenberg, G., Simmons, WH. Principles of
Medical Biochemistry, 3rd ed. Mosby Elsevier,
2012
 Devlin, TM. Textbook of Biochemistry, 6th ed.,
2006
 Koolman, J.; Roehm, KH. Color Atlas of
Biochemistry, 2 ed. Thieme, 2004