Plasma proteins and

Coagulation Factors
Abdul Salam M. Sofro
Faculty of Medicine
YARSI University Jakarta

Learning objectives
By the end of the lecture, students are
expected to understand :
various biochemical aspects of blood
plasma
Role and function of plasma factors in
hemostasis
Reference:
Murray K et al. 2000. Harpers Biochemistry, 25th ed.

Composition of Blood
Solid elements : RBC, WBC, Platelets
Liquid medium : plasma consisting of water,
electrolytes, metabolites, nutrients, proteins,
hormones, etc.
Water & electrolyte composition of
plasma is practically the same as that of all
extracellular fluids
Once the blood has clotted (coagulated),
the remaining liquid phase (called serum)
lacks of the clotting factors (including
fibrinogen)

Plasma proteins
Total plasma protein approx. 7.0-7.5 g/dl
A complex mixture of simple & conjugated
proteins such as glycoproteins & various types
of lipoproteins, thousands of antibodies
Can be separated
by sodium or ammonium sulfate into three major
groups fibrinogen, albumin & globulins
by electrophoresis using cellulose acetate into five
bands albumin, 1, 2, & globulin

Composition of blood plasma

Component
Water
Proteins
Salts
Lipids
Glucose (blood sugar)

Percent
~92
68
0.8
0.6
0.1

Serum
proteins

Cont.
Concentration of plasma protein is important
in determining the distribution of fluid
between blood & tissues
Osmotic pressure (oncotic pressure) exerted
by plasma protein is approx. 25 mm Hg.
Hydrostatic pressure in arterioles is approx. 37
mm Hg a net outward force of about 11 mm
Hg drives fluid out into interstitial spaces.
Hydrostatic pressure in venules is approx. 17
mm Hg a net force of about 9 mm Hg attracts
water back into circulation

Cont.
The above pressures are often referred to
as the Starling forces.
If plasma protein concentration is markedly
diminished (eg. due to severe protein
malnutrition fluid is not attracted back
into the intravascular compartment and
accumulates in extravascular tissue spaces
oedema

Cont.
Most plasma proteins are synthesized in the liver
Plasma proteins are generally synthesized on
membrane-bound polyribosomes
Almost all plasma proteins are glycoproteins
Many plasma proteins exhibit polymorphism
Each plasma protein has a characteristic half-life in
the circulation
The level of certain protein in plasma protein
increase during acute inflammatory states or
secondary to certain types of tissue damage

Some functions of plasma proteins

Antiprotease (antichymotrypsin, a1
antitrypsin, 2 macroglobulin, antithrombin)
Blood clotting (various coagulation factors,
fibrinogen)
Hormones
Immune defence (Ig, complement proteins,
2-microgloblin)
Involvement in inflammatory responses (acute
phase response protein eg. C-reactive
protein, 1-acid glycoprotein
Oncofetal (1-fetoprotein = AFP)

Cont.
Transport or binding proteins
albumin for bilirubin, FFA, ions, metals,
metheme, steroids, other hormones,
variety of drugs
Ceruloplasmin contains Cu but albumin
is more important in physiological transport
of Cu
Corticosteroid-binding globulin (transcortin)
Haptoglobin binds extracorpuscular Hb

Cont.
Liproproteins (chylomicron, VLDL, LDL,
HDL)
Hemopexin
Retinol-binding protein
Sex hormone-binding globulin
Thyroid-binding
Transferrin
Transthyretin (formerly pre-albumin, binds
T4 & forms a complex with Retinol-binding
protein)

Detail functions of some plasma

protein
Albumin:
Major protein of human plasma (3.4-4.7 g/dL)
Some 40% in plasma, 60% in extracellular space
Synthesized in liver as preproprotein, depressed in
a variety of diseases, particularly those of liver
(decreases albumin/globulin ratio)
Responsible for 75-80% of osmotic pressure of
human plasma
Ability to bind various ligands (include FFA, Ca,
certain steroid hormones, bilirubin etc.
Play an important role in transport of Cu, drugs

Cont.
Haptoglobin:
A plasma glycoprotein that binds
extracorpuscular Hb in a tight
noncovalent Hb-Hp complex
Prevent loss of free Hb into kidney
Its plasma levels are of some
diagnostic use low level in hemolytic
anemias

Cont.
Transferrin:
a 1-globulin, a glycoprotein, synthesized
in liver
Plays an important role in the bodys
metabolism of iron (two mol of Fe3+ per
mole of transferrin) diminishes potential
toxicity of free iron
Plasma concentration is approx. 300
mg/dL can bind 300 g of iron per dL
(Total Iron Binding Capacity of plasma)

 Ceruloplasmin (Cp)
2-globulin
Binds copper (Cu)
Exhibits a copper-dependent oxidase
activity
Low levels of Cp are associated with
Wilson disease
Tissue levels of Cu & certain other metals
are regulated in part by metallomethionins
(small protein found in the cytosol of cells
particularly liver, kidney & intestine)

 1-Antiproteinase (1-antitrypsin)
Synthesized by hepatocytes &
macrophages
Principal serine protease inhibitor of
human plasma inhibits trypsin,
elastase & certain other proteases
Deficiency of this protein has a role in
certain cases (approx. 5%) of
emphysema

 2-Macroglobulin
A large plasma glycoprotein
Comprises 8-10% of the total plasma
protein in human
Synthesized by a variety of cell types,
including monocytes, hepatocytes &
astrocytes.
Binds many proteinases (an important
panproteinase inhibitor)
Binds many cytokines

 Immunoglobulin
Play a major role in the bodys defence
mechanism
Synthesized by B lymphocytes

Immunoglobulin (Ig)
A group of proteins involved in mediating
immune response in higher organisms
In gamma globulin fraction of serum
Very heterogeneous
Similar in different species
106 different antibodies may be produced
in human adult

Ig structure
Tetramer :
* a pair of light chains (two identical
=kappa or =lambda chains)
* a pair of heavy chains (two identical
=alpha, =gamma, =delta, =epsilon or
=mu chains)
Light chain has one variable region (VL) &
one constant region (CL)
Heavy chain has one variable region (VH)
and three (, , ) or four (, ) constant
regions

Immunoglobulin
(antibody)
molecule

Ig class

Mol. Struct

Carbohydr

IgG

22

22

4 %

IgA

22

22

10 %

IgM

22

22

15 %

IgD

2 2

22

18 %

IgE

22

22

18 %

Ig functional groups
N terminal of H & L chains (VL/VH & CL /CH1) =>
antigen binding fragment
C terminal of L chain (CL) => interchain disulphide
bond
C terminal of H chain (CH) particularly C 2 & C 3
and C 4 of IgM & IgE) constitute the Fc fragment
responsible for class specific effector function =>
complement fixation or placental transfer, cell
surface binding etc

Schematic models of an antibody

molecule and a Fab fragment

Hemostasis and thrombosis

 Hemostasis is the cessation of bleeding from a

cut or severed vessel, whereas thrombosis

occurs when the endothelium lining blood
vessels is damaged or removed (eg. upon
rupture of an atherosclerotic plaque)
Hemostasis & thrombosis share three
phases:
Formation of a loose & temporary platelet
aggregate at the site of injury
Formation of fibrin mesh that binds to the
platelet aggregate, forming a more stable
hemostatic plug or thrombus
Partial or complete dissolution of the
hemostatic plug or thrombus by plasmin

Thrombi

Three types of thrombi:

White thrombus
Red thrombus
Disseminated fibrin deposit in very
small blood vessels or capillaries

Intrinsic and Extrinsic pathway of blood

coagulation
Two pathways lead to fibrin clot formation
These pathways are not independent
Initiation of fibrin clot in response to tissue
injury is carried out by extrinsic pathway,
but how intrinsic pathway is activated in
vivo is unclear (but it involves a negatively
charged surface)
Intrinsic & extrinsic pathways converge in a
final common pathway

 Involves many different proteins can be

classified into 5 types:
(1) zymogens of serine dependent
proteases which become activated
during the process of coagulation
(2) cofactors
(3) fibrinogen
(4) a transglutaminase, which stabilizes
fibrin clot
(5) regulatory & other proteins

Blood clotting factors

FI
: Fibrinogen
F II
: Prothrombin
F III
: Tissue factor
F IV
: Ca2+
FV
: Proaccelerin, labile factor,
accelerator (Ac-) globulin
F VII
: Proconvertin, serum prothrombin
conversion accelerator (SPCA),
cothromboplastin

Blood clotting factors

F VIII : Antihemophilic factor A,
antihemophilic globulin (AHG)
F IX : Antihemophilic factor B, Christmas
factor, plasma thromboplastin
component (PTC)
F X : Stuart Prower Factor
F XI : Plsm thromboplastin antecedent (PTA)
F XII : Hageman factor
F XIII : Fibrin stabilizing factor (FSF),
fibrinoligase

Intrinsic pathway
Involves factors XII, XI, IX, VIII, & X as
well as prekallikrein, HMW kininogen,
Ca2+ & platelet phospholipids results
in the production of factor Xa.
Commences with the contact phase in
which prekallikrein, HMW kininogen, F
XII & F XI are exposed to a negatively
charged activating surface.

Intrinsic pathway (cont.)

When the components of the contact
phase assemble on the activating surface,
F XII is activated to F XIIa upon proteolysis
by kallikrein. This F XIIa attacks
prekallikrein to generate more kallikrein,
setting up a reciprocal activation
F XIIa once formed, activates F XI to F XIa
and also release bradykinin from HMW
kininogen

Intrinsic pathway (cont.)

F XIa in the presence of Ca2+ activates F
IX. This in turn cleaves an Arg-Ile bond in
F X to produce F Xa

Intrinsic pathway
PK
HK
XII

XIIa
HK
XI

Ca 2+

Extrinsic pathway
VII

XIa
VIIa/Tissue factor
Ca 2+

IX
VIII

IXa
Ca 2+
PL

VIIIa

X
V

Xa
Va

Prothrombin

X
Ca 2+
PL
Thrombin

Prothrombin

Thrombin
XIII

Fibrinogen
XIIIa
Fibrin monomer

Fibrin polymer

Cross-linked
Fibrin polymer

Extrinsic pathway
Also leads to activation of F X but by
different mechanism.
Involves tissue factor, F VII, F X & Ca2+ and
results in the production of F Xa
It is initiated at the site of tissue injury with
the expression of tissue factor on
endothelial cells

Extrinsic pathway (cont.)

Tissue factor interacts with & activates F
VII. Tissue factor acts as a cofactor for F
VIIa, enhancing its enzymatic activity to
activate F X
Activation of F X provides an important
link between those two pathways

Final common pathway

Involves activation of prothrombin to thrombin
F Xa produced by either intrinsic or extrinsic
pathway, activates prothrombin (F II) to
thrombin (F IIa)
Activation of prothrombin, like that of factor X,
occur on the surface of activated platelets &
requires the assembly of a prothrombinase
complex, consisting of platelet anionic
phospholipid, Ca2+, F Va, F Xa, &
prothrombin

Final common pathway (cont.)

Conversion of fibrinogen to fibrin is
catalyzed by thrombin (thrombin also
converts F XIII to F XIIIa, a factor highly
specific transglutaminase that covalently
cross-links fibrin molecules by forming
peptide bonds between the amide groups
of glutamine & the e-amino groups of lysine
recidues, yielding a more stable fibrin clot
with increased resistance to proteolysis

Overview of hemostasis. a activated form of clotting factor V, VII,

VIII, IX, X; ADP, adenosine diphosphate; Epi, epinephrine; TXA2,
thromboxane; vWF, von Willebrand factor.

Some notes
Levels of circulating thrombin must be
carefully controlled achieved in 2 ways:
Feedback mechanism through a cascade
of enzymatic reactions for the conversion
of prothrombin to thrombin
Inactivation of any thrombin formed by
circulating inhibitors (the most important of
which is antithrombin III)

Some notes(cont.)
Endogenous activity of antithrombin III is
greatly potentiated by the presence of
heparin
Coumarin anticoagulants (eg. Warfarin)
inhibit vit.K-dependent carboxylation of F II,
VII. IX & X
Fibrin clots are dissolved by plasmin
(circulates in plasma in the form of its
inactive zymogen, plasminogen)

Some notes(cont.)
Activators of plasminogen are found in
most body tissues e.g.
tissue plasminogen activator (alteplase, tPA) is a serine protease that is released
into circulation from vascular endothelium
under condition of injury or stress & is
catalytically inactive unless bound to fibrin
(recombinant t-PA is used therapeutically
as a fibrinolytic agent as is Streptokinase
Urokinase (precursor: prourokinase)

Some notes(cont.)
Hemophilia A is due to deficiency of F VIII
Hemophilia B is due to deficiency of F IX
Endothelial cells synthesize prostacyclin
(potent inhibitor of platelet aggregation)&
other compounds that affect clotting &
thrombosis
Aspirin is an effective antiplatelet drug
Some laboratory tests measure coagulation
& thrombolysis