DBU,COLLEGE OF MEDICINE

Biochemistry of Hematology Module

By; Endalamaw Tesfa

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System Learning Objectives

At the end of this system students will be able to;
Explain about heme synthesis
Describe heme degradation
Describe the structures of hemoglobin & myoglobin
List the factors that affect oxygenation and
deoxygenation of hemoglobin
Explain about importance of plasma proteins

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Heme Synthesis
Iron is found in; Hgb, myoglobin, cytochromes,
catalase, peroxidase, hemosiderin, transferin, ferritin,
ferrochelatase, aconitase etc
The heme synthesized from glycine & succinyl-CoA
in the presence of Fe2+
Heme synthesized in almost all tissues in the body
which require hemoproteins.
Major sites of synthesis liver and bone marrow
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Inside RBCs, heme is synthesized in the normoblasts,

but not in the matured ones .
The heterocyclic porphyrin ring of heme consists of
four pyrrole rings
Heme is responsible for red color of the blood
Heme is the site at which each globin monomer binds
to one molecule of O2
Heme synthesis takes place in the mitochondria & in the
cytoplasm of the cell
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Heme Synthesis Pathway

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Heme
Synthesis
Pathway

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1. -Aminolevulinate (ALA) Synthase

Heme synthesis begins with condensation of glycine &
succinyl-CoA, with decarboxylation, to form aminolevulinic acid (ALA).
O

OOC

CH2

CH2

S-CoA +

succinyl-CoA
-Aminolevulinic
Acid Synthase

OOC

CH2

NH3+

glycine

H+
CoA-SH

CO2
O

OOC

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CH2

CH2

CH2

NH3+

-aminolevulinate (ALA)

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PLP serves as a coenzyme

ALA Synthase is the rate-limiting enzyme in heme
synthesis
Regulation occurs through control of gene
transcription.
Heme functions as a feedback inhibitor, repressing

transcription of the ALA Synthase gene in most cells.

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2. Porphobilinogen Synthase (PBG)

It is also called ALA Dehydratase, catalyzes
condensation of two molecules of -aminolevulinate to
form the pyrrole ring of PBG & release 2 H2O molecule.
COO

COO

CH2

CH2

CH2

CH2

+
O

PBG Synthase

2 H2O
O

CH2

CH2

NH3+

NH3+

2 -aminolevulinate
(ALA)
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H2C

COO

COO

CH2

CH2

CH2

CH

NH3+

N
H

porphobilinogen
(PBG)

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3. Porphobilinogen Deaminase (PBG)

COO

N
H

pyrrole

COO

CH2

CH2

CH2

H2 C
NH3+

N
H

Porphobilinogen (PBG)

Porphobilinogen (PBG) is the first pathway

intermediate that includes a pyrrole ring.
The porphyrin ring is formed by condensation of 4
molecules of porphobilinogen.
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Porphobilinogen Deaminase catalyzes successive

PBG condensations, initiated in each case by

elimination of the amino group.

Porphobilinogen Deaminase has a dipyrromethane
prosthetic group, linked at the active site via a cysteine S.
The enzyme itself catalyzes formation of this prosthetic
COO-

group.
COO

CH2

Enz

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COO

CH2

COO

CH2

CH2

N
H

dipyrromethane

CH2
CH2

N
H

PBG Deaminase

PDB
1PDA
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PBG units are added to the dipyrromethane until a

linear hexapyrrole has been formed.
Porphobilinogen Deaminase is organized in 3 domains.
Predicted interdomain flexibility may accommodate
the growing polypyrrole in the active site cleft.
Hydrolysis of the link to the enzyme's dipyrromethane
releases the tetrapyrrole hydroxymethylbilane.

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Linear Structure of Hexapyrrole

COO COO -

CH2
COO - COO -CH

COO -

Enz

COO -

CH2

COO -

CH2

CH2

CH2

CH2

CH2

N
H

CH2

CH2
CH2

N
H

NH

HN

NH

HN

CH2 COO -

CH2 COO CH2

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CH2 CH2

CH2

COO -COO -

CH2
COO -

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4. Uroporphyrinogen III Synthase

Uroporphyrinogen III Synthase converts the linear
tetrapyrrole hydroxymethylbilane to the macrocyclic
uroporphyrinogen III.
COO -

hydroxymethylbilane

OOC

COO -

CH 2

COO -

CH 2

COO

CH 2

CH 2

CH 2

CH 2

CH 2

CH 2
NH

HN

NH

HN

COO -

CH 2

OOC

uroporphyrinogen
III
-

CH 2
NH

HN

NH

HN

CH 2

CH 2

CH 2

COO -

COO -

HO
C

C
CH 2

COO

OOC CH 2

CH 2

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CH 2 CH 2

CH 2

COO -COO -

CH 2

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COO -

Uroporphyrinogen III
Synthase

CH 2

CH 2

CH 2

CH 2

COO -

COO -

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Uroporphyrinogen III Synthase catalyzes ring closure

& flipping over of one pyrrole to yield an asymmetric

tetrapyrrole.
This rearrangement is thought to proceed via a spiro
intermediate.
Note the distribution of acetyl & propionyl side chains,
as flipping over of one pyrrole yields an asymmetric
tetrapyrrole.

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The active site of

Uroporphyrinogen III
Synthase is in a cleft
Uroporphyrinogen III
PDB 1JR2
Synthase

between two domains of the

enzyme.

The structural flexibility inherent in this arrangement is proposed

to be essential to catalysis.
Uroporphyrinogen III is the precursor for synthesis of vitamin B12,
chlorophyll, and heme, in organisms that produce these compounds.

Conversion of uroporphyrinogen III to protoporphyrin IX occurs

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in several
steps.

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5. Uroporphyrinogen Decarboxylase
All 4 acetyl side chains are decarboxylated to methyl
groups by Uroporphyrinogen Decarboxylase
Oxidative decarboxylation converts 2 of 4 propionyl
side chains to vinyl groups by Coproporphyrinogen
Oxidase
Oxidation adds double bonds by Protoporphyrinogen
Oxidase
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Uroporphyrinogen Decarboxylase
COO

CH2

COO

CH2

CH2

CH2

CH

OOC CH2

CH2

CH2 COO-

CH3

CH CH2

H3C

NH

HN

NH

NH

HN

CH2

OOC CH2

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protoporphyrin IX

uroporphyrinogen III

COO

HN

H3C

CH3

CH2

CH2

CH2

CH2

CH2

CH2

CH2

CH2

COO-

COO

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COO

COO

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6. Ferrocheletase
CH2

protoporphyrin IX

CH2

CH

CH3

CH

CH CH2

H3C
NH

Fe

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CH2

CH2

CH2

CH2

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CH CH2

COO

2H

Fe
N

H3C

Ferrochelatase

N
CH3

CH3

H3C

HN

H3C

COO

++

heme

CH3

CH2

CH2

CH2

CH2

COO

COO

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Fe++ is added to protoporphyrin IX via

Ferrocheletase, a homodimeric enzyme containing 2 iron-

sulfur clusters.
A conserved active site His, along with a chain of
anionic residues, may conduct released protons away, as
Fe++ binds from the other side of the porphyrin ring, to

yield heme.

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Regulation of Heme Metabolism

In erythrocytes and hepatocytes, ALA synthase is the
rate controlling enzyme.
There are two forms of ALA synthase
ALA synthase 1 is downregulated by heme
ALA synthase 2 is downregulated by insufficient iron
Under physiologic conditions sufficient iron upregulated
ALA synthase 2 in the developing erythrocytes.

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In contrast, under physiologic conditions in liver cells,

ALA synthase 1 is present in relatively low amount
ALA synthase 1 in the liver has a short life span, as
does the mRNA coding for it.
Agents that alter the rate of synthesis of the mRNA for
ALA synthase 1 have a rapid effect on the amount of ALA
synthase 1 and in the synthesis of porphyrins and heme.

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Porphyrias:
It is a group of diseases, caused by a specific abnormality
in the heme synthesis pathway
Porphyria is most often an inherited mutation in one of
the genes involved in heme production, & some
environmental factors.
Hgb is a protein in RBCs that carries O2
Heme also plays a role in breaking down chemicals that
removed from the body
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Heme is synthesized mainly in the bone marrow & liver

Eight different enzymes are involved in heme synthesis
Deficiency of a specific enzyme in heme synthesis can
result in increase in porphyrins, and deposited in the skin,
in the bone, in the teeth and causes different symptoms.
The eight type of porphyria is occur due to the
deficiency of different enzyme.
Porphyria classified based on their site of defect,
acuteness or chronicness
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Acute Porphyria :
Aminolevulinate dehydratase deficiency porphyria (ALA-D)
Acute intermittent porphyria (AIP)
Hereditary coproporphyria (HCP)
Variegate porphyria (VP)

Chronic Porphyria:
Porphyria cutanea tarda (PCT)
Erythropoietic protoporphyria (EPP)
Congenital erythropoietic porphyria (CEP)

Hepatoerythropoietic porphyria (HEP)

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Aminolevulinate Dehydratase Deficiency

Porphyria (ALA-D)
-aminolevulinate dehydratase (ALAD) deficiency
porphyria is an autosomal recessive disorder caused by a
homozygous ALAD deficiency
ALAD is the second enzyme in the heme biosynthesis
This enzyme activity is present in large excess
compared to other enzymes in the heme synthesis

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Acute Intermittent Porphyria

Its is a hereditary autosomal dominant occur due to
the deficiency of hepatic uroporphyrinogen I synthase
Porphobilinogen accumulates, & is excreted in the
urine.
Heme synthesis is reduced.
ALA synthase activity therefore increases.

There are neurological symptoms, which cannot be

explained.
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Hereditary Coproporphyria (HCP)

HCP is an inherited autosomal dominant disorder
HCP results from a deficiency of coproporphyrinogen
oxidase (CPO).
Many molecular defects in CPO give rise to HCP, with a
variable phenotype.
Many HCP patients have increased excretion of
coproporphyrin III in the feces & urine.
Some patients have had jaundice & hepatic dysfunction.
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Variegate porphyria (VP)

VP is an inherited autosomal dominant disorder
VP usually results from a heterozygous deficiency of
~50% protoporphyrinogen oxidase (PPO) activity.
The presentation of VP is variable, hence its name
Variegate porphyria
VP is characterized by increased fecal protoporphyrin &
also coproporphyrin.
In the urine there is increased ALA, PBG, &
coproporphyrin.
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Porphyria Cutanea Tarda (PCT)

PCT is a group of acquired and familial disorders
deficiency of uroporphyrinogen decarboxylase
(UROD) enzyme
Approximately 80% of all cases of PCT are acquired;
20% are familial, but it vary among different geographic
regions & ethnic groups.
Familial PCT occur single gene mutation of UROD
Hepatic tumors producing excess porphyrins are rare
causes
ofT PCT
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Erythopoietic Protoporphyria (EPP)

EPP is the commonest of the erythropoietic porphyrias
and second only to PCT in prevalence of all the porphyrias
It equally affects both sex and all ethnic groups.
The clinical expression is highly variable.
Photosensitivity is the major clinical manifestation
Cutaneous symptoms like; burning, itching, or pain in
the skin on exposure to sunlight
It is variable among patients even of the same family
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Congenital Erythropoietic Porphyria (CEP)

CEP is a rare condition resulting from an inborn error in
prophyrin metabolism.
Due to defect of uroporphyrinogen III synthase .
Deficiency of UROS results in excess uroporphyrin I, &
coproporphyrin I, accumulated in bones, erythrocytes, skin,
& teeth which causes photosensitization, severe anaemia
CEP patients treated by bone marrow transplantation

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Hepatoerythropoietic Porphyria (HEP)

HEP is a rare form of porphyria caused by deficiency of
uroporphyrinogen decarboxylase (UROD) enzyme
Excess porphyrins synthesized in the liver & bone marrow.
The clinical diagnosis is based on elevated urinary uro- and
heptacarboxyl-porphyrins & zinc protoporphyrin in
erythrocytes is elevated.
Several mutations and deletions have been described.
The prognosis of HEP is poor due to the severe defect in

UROD activity
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Heme Degradation
Most of the heme obtained from Hgb in RBCs
100 200 million aged RBCs are broken down /hour
There is a turnover of about 6 g/day of Hgb
Senescent RBCs and heme from other sources are
engulfed by cells of the RES.
The globin is recycled or converted into AAs
Heme is oxidized.

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1. Conversion of Heme to Bilirubin

It takes place in the RES
Heme ring is cleaved by heme oxygenase
Biliverdin reductase reduces the central methene
bridge of biliverdin, producing bilirubin.
The high lipid solubility of bilirubin determines its
behavior and its further metabolism.
It transported in the blood by serum albumin.
It is soluble in the lipid bilayers of cell membranes.
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Conversion of Heme to Bilirubin

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2. Bilirubin Conjugation with Glucuronic Acid:

Conjugation of bilirubin made bilirubin water soluble
Conjugation is accomplished by attaching two
molecules of glucuronic acid by UDP glucuronyl
transferase
The reaction is a transfer of two glucuronic acid
groups sequentially to the propionic acid groups of the
bilirubin.
The major product is bilirubin diglucuronide.
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Bilirubin
diglucuronide is excreted in the bile.
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 It is subject to subsequent transformations to other

species by the intestinal bacteria

In small intestine bilirubin changed to urobilinogen by

intestinal bacteria
Some urobilinogen (UBG) reabsorbed by intestinal
mucosa and the rest excreted in urine or oxidized to
urobilin and excreted in the feces

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Bilirubin Diglucuronide

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3. Metabolism of CB by Intestinal Bacteria

In normal individuals, intestinal bilirubin is acted on by
bacteria to produce the final porphyrin products,

urobilinogens and urobilins, that are found in the feces.

A small fraction of urobilinogen is reabsorbed into the
blood, extracted by the kidney, and excreted in the urine.
Bile pigments are bilirubin and its catabolic products

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Hyperbilirubinemia (Jaundice)

1. Pre-hepatic
It is caused by increased hemolysis and increased
degradation of heme
Patients with sickle cell anemia and other hemolytic
diseases
Pre-hepatic jaundice is indicated by increased
unconjugated bilirubin with relatively normal serum
conjugated bilirubin and increased urinary urobilinogen.
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2. Hepatic Hyperbilirubinemia
It is generally due to defective conjugation of bilirubin
by hepatocytes resulting in increased serum
unconjugated and conjugated bilirubin
Causes include the genetic disorders like Gilberts and
Crigler-Najjar syndromes; Neonatal jaundice, and viral
and chemical damage to hepatocytes
The rare congenital biliary atresia with abnormal
anatomic obstruction of biliary ducts could be another
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2.1 Gilbert's disease

May be caused by an inability of the hepatocytes to

uptake bilirubin from the blood

As a result, unconjugated bilirubin accumulates.

2.2 Physiological jaundice & Crigler-Najjar syndrome

Conjugation is impaired.
Unconjugated bilirubin is retained by the body.

2.3 Dubin-Johnson syndrome

Inability of the hepatocytes to secrete conjugated bilirubin
after it has been formed.
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Conjugated
bilirubin returns to the blood.

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3. Post-hepatic (Biliary obstruction)

It is generally due to a defect in transporting
conjugated bilirubin and bile out of the liver
It can involve obstruction of the small canaliculi within
the liver, the hepatic bile duct, and the common bile duct
Obstruction of the bile flow can be due to cirrhosis,
gallstones or tumors
Blood levels of conjugated bilirubin increase

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Hgb Structure & Function

Hgb is tetramer protein found in RBCs
It is used to transport 4O2 from the lungs to the tissues
& CO2 from the tissues to the lungs.
Adult Hgb, is tetramer 2 & 2 chains held by noncovalent interactions.
Each subunit has stretches of -helical structure, and a
heme-binding pocket similar to that of myoglobin.
Hgb molecule is more complex than myoglobin.
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The binding of O2 to the heme iron pulls the iron into

the plane of the heme
The deoxy Hgb is called T, or taut form.
The binding of O2 to Hgb causes the rupture of some
of the ionic bonds & hydrogen bonds between the
dimers forms R, Hgb
The R form is the high O2 affinity form of Hgb.

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R and T states
The distance between two Fe atoms of the chains
decreases from 40 to 33 oA on oxygenation.
When oxygenation takes place, large structural changes
take place at the 1 2; 2 1 contact but only small
changes occur at 11; 2 2 contact.
Also the 11; 2 2 contact pair rotates 15 degrees
relative to the other pair, this involves breakage of the salt
bridges at the contact points.
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The change in 1 2 ; 2 1 contact region acts as a

switch between the two structures.

The valine residue in the heme pocket moves, opening

the cleft for oxygen binding.
The central pocket of the tetramer becomes too narrow
to accommodate 2,3-BPG, which is expelled out.

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Transition from T to R state

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Oxygen Dissociation Curves

The binding properties of O2 transport proteins are
described by plotting their percent saturation against pO2:
O2 dissociation curves.
When O2 binds to the first subunit of deoxyhemoglobin
it increases the affinity of the remaining subunits for O2.
O2 dissociation curves of Myoglobin: Hyperbolic
O2 dissociation curves of Hgb: Sigmoidal
At 100 torre pO2, Hgb is fully saturated with O2.
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Oxygen Binding Curves of Mb & Hb

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As oxyhemoglobin circulates to deoxygenated tissues,

O2 is unloaded and the affinity of Hgb for O2 is reduced

This facilitating the unloading of next O2 molecules.

Low O2 tension is found in very active exercising
tissues, the binding affinity of Hgb for O2 is very low
allowing maximal delivery of O2 to the tissue.

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O2 Dissociation Curve:
The standard curve is shifted to the right by an increase
in temperature, 2,3-BPG, or pCO2, or a decrease in pH.
The curve is shifted to the left by the opposite of these
conditions.
A rightward shift, by definition, causes a decrease in
the affinity of Hgb for O2.
Rightward shift makes harder for the Hgb to bind to O2,
but it makes easier for the Hgb to release bound O2.
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Leftward shift increases the affinity, making the O2

easier for the Hgb to pick up but harder to release.

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Factors Affecting O2 Dissociation Curve:

The effectiveness of hemoglobin-oxygen binding can be
affected by several factors;
1. Increased temperature (e.g., exercise).
2. Increased [H+] or pH
3. Increased 2,3-BPG
4. Increased CO2
5. Carbon monoxide (CO)

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1. Temperature
Increases O2 unloading in working muscle
Temperature does not have a dramatic effect as other
factors
Hyperthermia causes a rightward shift
Hypothermia causes a leftward shift

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2. Bohr Effect
Decreased pH leads to decreased ability for Hgb to
hold O2 /Hgb increased ability to leave O2
Bohr effect is seen in exercising muscle where
anaerobic metabolism results in lower pH.
Bohr effect helps with the offloading of O2 from.
The acidosis that accompanies tissue hypoxia from
whatever cause improves O2 release in metabolically
challenged areas.
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Bohr effect helps to increase fetal oxygenation.

The transfer of CO2 from fetal to maternal blood shifts
the maternal oxyhaemoglobin curve to the right and the
fetal curve to the left
Facilitating the transfer of O2 across the placenta from
mother to fetus.

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Chloride Shift
Movement of chloride ions into RBCs in exchange to HCO3During isohydric transport of CO2, HCO3- diffuses out of
RBCs.
To maintain electrical neutrality in RBCs, an anion must
move into the cells.
The formation and ionization of carbonic acid in plasma is a
major mechanism for the transport of CO2 to the lungs, i.e. in
the plasma as HCO3-.

H+ produced in plasma by the ionization of carbonic acid is

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buffered by phosphate (HPO42-) and by proteins.

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3. Effect of 2,3-BPG on O2 Affinity:

2,3-BPG is an important regulator of the binding of O2
to Hgb.
It is the most abundant organic phosphate in the RBC
2,3-BPG synthesized from an intermediate of glycolysis
2,3-BPG decreases the O2 affinity of Hgb by binding to
deoxy - hemoglobin but not to oxyhemoglobin.
This preferential binding stabilizes the taut
conformation of deoxyhemoglobin.
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Binding site of 2,3-BPG:

One molecule of 2,3-BPG binds to a pocket, formed
by the 2 -globin chains, in the center of the
deoxyhemoglobin tetramer.
This pocket contains several positively charged AAs
that form ionic bonds with the negatively charged
phosphate groups of 2,3-BPG.
2,3-BPG is expelled on oxygenation of the Hgb.

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Shift of O2 Dissociation Curve:

Hgb from which 2,3-BPG has been removed has a
high affinity for O2.
In RBC, the presence of 2,3-BPG significantly
reduces the affinity of Hgb for O2, shifting the O2
dissociation curve to the right.
This reduced affinity of Hgb to O2, it release O2
efficiently at the partial pressures found in the tissues.

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Effect of BPG in O2 Delivery;

Conversion of R to T state upon BPG binding facilitates
O2 delivery by bringing about a right shift in O2 dissociation

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4. Binding of CO2:
Most of the CO2 produced in metabolism is transported
as bicarbonate ion.
Some CO2 is carried in the form of
carbaminohemoglobin.
The binding of CO2 stabilizes the T form of
hemoglobin, resulting in a decrease in its affinity for
oxygen and a right shift in the oxygen dissociation.
In the lungs, CO2 dissociates from the Hgb, & is
released
inT the breath.
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5. Binding of CO:
CO binds tightly to the Hb iron, forming
carboxyhemoglobin and prevents O2 binding.
When CO binds to one or more heme sites, Hgb shifts
to the relaxed conformation, causing the remaining heme
sites to bind O2 with high affinity.
This shifts the O2 dissociation curve to the left, &
changes the normal sigmoidal shape toward a hyperbola.
The affected Hgb unable to release O2 to the tissues.
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Effect of CO poisoning on Hgb Dissociation curve

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Myoglobin
It is Monomeric heme protein made up of 153AAs
It is mainly found in the cytoplasm muscle tissue &
serves as intracellular reservoir of O2.
Myoglobin binds very tightly with O2 & releases it only
at very low pO2
The released O2 can then be utilized for the metabolic
activity of muscle cells.
The myoglobin curve is a rectangular hyperbola with a
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very
low
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T P50 (2.75 mmHg).

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It lies well to the left of the sigmoid-shaped Hgb curve.

It has much higher oxygen affinity.

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Physiological Importance of Myoglobin

Myoglobin in tissues accepts O2 from Hgb in
circulating arterial blood, then delivers it to the
mitochondria when O2 needs are sufficiently great.
The p50 for myoglobin is very low, signifying that
myoglobin has a high affinity for O2
Myoglobin extract O2 from Hgb
At the O2 concentration existing in tissues, myoglobin is
nearly saturated.
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As the cell carries out its metabolic activities, it utilizes O2 &

the internal pO2 falls to levels where myoglobin releases its O2.

Still myoglobin p50 (1 mm Hg) is too low to release any

significant amount of O2 in normal resting tissues;
Rapidly exercising muscle or hypoxia would myoglobin
release its O2
Due to its hyperbolic curve & very low p50, myoglobin is
unsuitable for the transport of O2 as it is insensitive to small
changes pO2.
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Biochemical Reason:
Both of them had different structures
Myoglobin is monomeric protein & its dissociation
curve is rectangular hyperbola.
Hgb contains four globin chains & the oxygenation of
one chain shows positive cooperativity .
This cause of the sigmoid shape of the dissociation
curve.

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Plasma Proteins
Plasma is clear creamy/yellowish cell free fluid
Cells contribute about 40 50% of the blood volume.
Plasma carries most of the dissolved nutrients &
metabolites & helps in their distribution and transport
between the tissues and body compartments.
Hundreds of proteins circulating in plasma perform
diverse functions like hormonal, catalytic, transport,
defence, blood clotting, anticoagulants and fibrinolysis
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Functions of Plasma Proteins

Intravascular osmotic effect
Viscosity
Transport of insoluble substances
Protein Reserve
Clotting
Inflammatory Response
Protection from infection
Maintenance of acid-base balance
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Composition of Human Plasma

Electrolyte/ Gas Normal (adult)
Levels[mmol/L]
CO2 (Arterial)

21 28

CO2 (venous)

24 30

Bicarbonate

21 28

Chloride

95 103

Sodium

136 142

Potassium

3.8 5.0

Calcium

2.3 2.74

Magnesium

0.65 1.23

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Metabolite
Glu (Fasting)
Total Lipids
Total Proteins
Urea
Uric acid
Creatinine
Bilirubin
Bile acids
Acetone
Acetoacetic
acid

Normal (adult)
70 100 mg/dL
400 800 mg/dL
6.0 - 8.0 g/dL
20 40 mg/dL
2.7 7.0 mg/dL
0.6 1.2 mg/dL
0.1 1.2 mg/dL
0.3 3.0 mg/dL
0.3 2.0 mg/dL
0.2 1.0 mg/dL
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Turnover of Plasma Proteins

Most plasma proteins synthesized by liver
50 gm proteins synthesized by the liver everyday
Immunoglobulins: only proteins that are not synthesized
in liver rather they produced by plasma cells.
All plasma proteins, except albumin, are glycoproteins
and carry an oligosaccharide moiety.
The receptor mediated endocytosis of glycoproteins
leads to their removal from circulation and degradation in
lysosomes.
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Albumin
Synthesized in hepatic parenchyma cells
Liver produces about 12 g albumin every day.
Most abundant (~4.5 g/dl) of plasma proteins
Half life: 15-19 days.
Single polypeptide chain of 585 AAs; MW: 66.3 kD.
Due to small size, it can seep out of the blood vessels & is a
component of most extravascular fluids e.g. CSF, interstitital

fluid, urine & amniotic fluid.

About 60% of the total body albumin is extra-vascular.
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Functions of Albumin
Maintenance of osmotic pressure
Transports FFA, PLs, metal ions, AAs, drugs,
hormones, bilirubin.
Reservoir for many hormones & metabolites.
Binding kinetics of drugs with albumin is important
determinant for the dose and duration of action of drugs.
AA source for peripheral tissues.
Liver stores extra AAs after meals into albumin
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Transport of AAs across membranes e.g. Trp is bound to

albumin for transport across BBB
Component of plasma anti-oxidant activity as it binds
with FFAs, divalent ions, hypochloride, bilirubin.
Although isoeletric pH of albumin is on the acidic side,
its ability to bind with free ions and other charged
molecules is increased when the blood pH deviates away
from physiological pH.

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Clinical Significance of Albumin

Serum Albumin due to;
An inadequate source of amino acids that occurs in
malnutrition and malabsorption
Liver disease, resulting in decreased synthesis by the
hepatocytes.
Protein-losing enteropathy or gastrointestinal loss as
interstitial fluid leaks out in inflammation and disease of the
intestinal tract as in diarrhea

Kidney
loss
to
the
urine
in
renal
disease
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Skin loss in the case of burns or exfoliative dermatitis

Hypothyroidism

Dilution by excess fluid

Acute disease states
Mutation resulting from an autosomal recessive trait
Redistribution by hemodilution, increased capillary
permeability, or decreased lymph clearance.

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Acute Phase Reactants:1-Acid Glycoprotein

Synthesized primarily in liver
Monomer, 181 AAs, MW 40 kD
Plasma half life: about 3 days.
It binds with a number of lipophilic substances.

Inhibits mitosis of viruses /parasites control

replication
Inhibits platelet aggregation & phagocytosis by
neutrophils.
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Significance of 1-Acid Glycoprotein

Increased plasma levels during Inflammation& necrosis
with levels peaking 3 to 5 days after initial exposure.
Glucocorticoids stimulate the synthesis of AAG,
therefore, steroid hormonal administration or even
endogenous over-synthesis result in increased levels of
AAG.
Decreased levels of AAG are seen in nephrotic
syndrome, protein losing enteropathy and in conditions
associated
with increased estrogens.
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Relative increase in the acute phase reactants after

infection or trauma.

CRP : C-Reactive Protein; AAG : 1-Acid Globulin;

AAT : 1-Anti-Trypsin; C3 : Complement C3 protein

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C-Reactive Protein
Synthesized in liver
CRP bind with C-polysaccharide on the cell wall of
Streptococcus pneumoni bacteria, fungi, protozoa.
Binding activates the classical complement pathway to
initiate opsonization, phagocytosis & lysis of the
antigens.
Detoxification of endogenous toxic substances
released from damaged tissues by binding with them
leading to their clearance from the blood.
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Clinical Significance of CRP

CRP raised within 24 to 48 hours in MI, stress, trauma,
infection, surgery or malignancy up to 2000 times.
Estimation of CRP in plasma is used to:
Assess inflammatory activity in rheumatoid arthritis.
Detect infections in systemic lupus erythromatosus
(SLE), leukemia or after surgery.
Detect rejection of the transplanted kidney or other
tissues.
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Manage
neonatal septicemia and meningitis.
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1-Antitrypsin(AAT)
AAT is a very small molecule and can easily diffuse
into the interstitial fluids /tissues & protects them
against protease digestion.
Single polypeptide 394 amino acids
Carries three oligosaccharide chains.
Primarily synthesized in liver, also locally contributed
by monocytes and macrophages.
Plasma half life: 6 to 7 days.
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Function of 1-Antitrypsin(AAT)
Most important inhibitor of leukocyte elastase, released
during phagocytosis by the polymorphonuclear cells.
Inhibitor of leukocyte neutral protease, kallikrein,
rennin, urokinase, plasmin and thrombin as well as other
enzymes of the clotting cascade .
Acute phase reactant Its concentrations are
significantly increased during inflammatory and necrotic
processes.
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Clinical Significance of 1-Antitrypsin(AAT)

Increased AAT levels:
Seen in acute phase reaction .
Uptake of AAT-protease complex by hepatocytes

Observed in cases with inflammation of hepatic

parenchyma i.e. hepatitis.
Estrogen stimulates AAT synthesis, therefore, during
late pregnancy as well as during estrogen therapy, AAT
levels could be raised.
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Decreased AAT levels:

Genetic deficiency of AAT leads to pulmonary emphysema .
AAT deficiency is also associated with neonatal hepatitis,
cirrhosis and liver carcinoma.
In neonatal respiratory syndrome, AAT is utilized at a faster
rate leading to its lower plasma levels.
AAT is a very small molecule and hence is easily filtered
through glomerulus.

In nephritic syndrome or in renal tubular disease, the plasma

AAT could be decreased due to urinary loss of AAT.
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2-Macroglobulin (AMG)
Protease inhibitor of plasma.
Synthesized by liver parenchyma cells
2-Macroglobulin is primarily an intravascular
protein and is found in CSF or urine only under
pathological conditions like meningitis and renal
failure, respectively.
Major 2-globulin of plasma and contributes about 8
to 10% to the total plasma proteins.
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Functions of 2-Macroglobulin (AMG)

Transport of a number of small peptides such as
cytokines and growth factors.
Binding of cytokines to AMG reduces their stimulation
of the synthesis of acute phase reactants, thus controlling
the acute phase reactions and inflammation.
Transport of divalent ions particularly Zinc.
About 10% of the zinc in plasma is transported bound
to AMG.
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Clinical Significance of 2-Macroglobulin

Increased levels:
Seen in females on estrogen therapy & in nephritic
syndrome.
Infants and children have higher plasma concentrations
than adults.

Decreased levels:
Observed in acute phase reaction, pancreatitis and
pancreatic carcinoma.
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Prothrombin
Single polypeptide 2-globulin
During injury, prothrombin is activated to thrombin by
activated factor Xa by the cleavage of an N-terminal
peptide.
Thrombin is a serine protease with MW of 34 kD.
Thrombin gets attached, through negative charges on
the surface of platelets.
Calcium helps in its interaction with the platelets.
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Fibrinogen
Fibrinogen is a dimeric molecule that is composed of two
sets of three polypeptide chains termed A, B, and .
Fibrinogen and fibrin play important roles in blood
clotting, fibrinolysis, cellular and matrix interactions,
inflammation, wound healing, and neoplasia.
The polypeptides are joined by disulfide bridging within
the N-terminal E domain.

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Plasmin and Fibrinolysis

The fibrin clot must be dissolved after the injury healed.
Thrombosis is a dangerous condition and puts the person
at risk of coronary artery disease or stroke.
The enzyme responsible for dissolution of the clot is
plasmin.
Plasmin exists in blood as zymogen plasminogen,
which is converted to active plasmin by the proteolytic
action of tissue-specific or urokinase-type plasminogen
activators.
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Transferrin( TRF)
Single polypeptide iron transport protein in plasma
Synthesised in liver and to a small extent in
reticuloendothelial cells, testes and ovaries
Plasma half life: About 7 days.
About 50% is present in the extravascular compartment.
Transferrin has two binding domains for polycations and

has an affinity for binding to iron (Fe3+), copper, zinc,

cobalt & calcium.
Plasma levels of TRF are regulated by the plasma iron
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Function of Transferrin
It transfers iron (Fe3+) to liver & reticuloendothelial
system.
Apo-tranferrin binds iron absorbed from intestine or
released from Hgb, in ferric form.
Most body cells have receptors for transferrin the
receptors have higher affinity for TRF-Fe3+ complex
than Apo-TRF.
Once bound, the complex is internalized through coated
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pits
which
form endosomes .
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Inside the endosomes, low pH triggers the release of

iron from TRF.

The released iron diffuses out into the cell and is

incorporated into ferritin and hemosiderin.
The stored iron then can be used for the synthesis of
Hgb, myoglobin and cytochromes.
The apo-TRF returns to the surface and is released
into plasma.

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Thanks

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