Struktur, Fungsi dan

Metabolisme Eritrosit Dan

Lekosit

Dr. Marhaen Hardjo, M.Biomed,

PhD
Bagian Biokimia Fakultas Kedokteran
Universitas Hasanuddin Makassar

The Structure and

Function
of Blood

Composition of Blood

Blood is responsible for..

Transporting gases (oxygen & carbon

dioxide)
Transporting waste products
Transporting nutrients
Helping remove toxins from the body

Composition of Blood

Blood makes up 68% of our

total body weight.

Normal adult blood volume is 5 L.

Blood is made up of cellular

material in a fluid called plasma.

Composition of Blood

Blood is a circulating tissue

consisting of three types of cells.
1.
2.
3.

Red Blood Cells Erythrocytes

White Blood Cells Leukocytes
Platelets Thrombocytes
The cells listed above are suspended in
a liquid known as plasma.

Formation of Blood
Hematopoiesis the formation and development of blood cells
In adults the cellular elements are produced in the bone marrow.
Some WBCs are produced in the lymphatic tissue and bone
marrow.
Blood cells need certain nutrients to form properly.
Examples include..
Iron
Folic acid
Vitamin B12
All blood cells formed come
from a hematopoietic stem cell.

These cells can become any

blood cell.

Composition of Blood

The blood is made up of cells

that are suspended in liquid
called plasma.

Plasma makes up 55% of the blood.

Plasma is made of 90% water and

10% proteins, lipids, carbohydrates,
amino acids, antibodies, hormones,
electrolytes, waste, salts, and ions

Blood cells make up the remaining

45% of the blood.

Red blood cells make up 99% of the blood cells.

White blood cells and platelets make up the other 1%.

Composition of Blood

Each type of blood cell performs a different function.

Red blood cells (Erythrocytes)

White blood cells (Leukocytes)

Platelets (Thrombocytes)

Composition of Blood:
Red Blood Cells

Red Blood Cells

AKA: Erythrocytes or RBCs

Most abundant cell in the blood

(4 million 6 million per microliter of blood)

Formed in the bone marrow

Main function is transporting

oxygen and carbon dioxide

Mature forms do NOT have a nucleus

Shaped as biconcave disks

6-8 micrometers in diameter

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Composition of Blood:
Red Blood Cells

Red Blood Cells

Stain pink-tan

Center of cell is lighter

central area of pallor

Life span of about 120 days

Hemoglobin (iron protein)is

found in the RBC

Hemoglobin carries oxygen from the

lungs to the rest of the body and carbon
dioxide binds to the RBC and is taken to
the lungs to be exhaled.

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Metabolisme Sel Darah

Merah

Sel darah merah yang masih muda

mempunyai metabolisme yang sangat
aktif karena organel selnya masih lengkap
Sel darah merah dewasa tidak lagi
mempunyai aktivitas metabolisme seperti
sel muda tersebut
Tetapi Sel darah merah dewasa masih
memerlukan energi untuk menjalankan
fungsinya
11

2 jenis metabolisme pada

sel darah merah dewasa:

Glikolisis
Fosfoglukonat (HMP-shunt)

12

Overview Carbohydrates
Metabolisme
Glucose

Hexokinase
Pentose
Phosphate
Shunt

Glucose-6-P
glycolysis

Pyruvate

Glc-1- phosphate

glycogen
13

cytosol

Pyruvate

mitochondria
(aerobic)

FATTY ACIDS

AMINO
ACIDS

Acetyl CoA

Krebs
cycle

Reducing
equivalen

Oxidative
Phosphorylati
(ATP)

14

Glikolisis

jalur metabolisme Embden-Meyerhof.

jalur oksidasi glukosa menjadi
piruvat +
energi (ATP) + NADH.
berlangsung dalam sitosol.
jalur sumber energi terutama bagi
sel darah merah dewasa yang tidak lagi
punya mitokondria.
15

PENGERTIAN
GLIKOLISIS
@.PEMECAHAN GLUKOSA=OKSIDASI GLUKOSA
@.C6H12O6
CO2 + H2O + 38 ATP
@. EMBDEN MEYERHOFF: GLUKOSA/PIRUVAT
8 ATP ( AEROB).HANYA 2 ATP BILA ANAEROB
@. PIRUVAT / ASETIL KoA : 2NADH=6ATP
@. KREBS: 2X12=24 ATP
TOTAL
38 ATP
BILA ANAEROB: 2 ATP + LAKTAT
16

Nasib Piruvat

Piruvat dapat mengalami proses:

1. Aerobik (ada oksigen),
piruvat dioksidasi untuk menghasilkan
CO2 dan H2O melalui daur Krebs
2. Anaerobik (tanpa oksigen),
manusia = piruvat laktat
ragi
= piruvat etanol
27

Lactate Dehydrogenase
O

NADH + H NAD

C O

C
HC OH

CH3

CH3

pyruvate

lactate
28

29

Pengaturan Glikolisis
hexokinase or
glucokinase
phosphofructokinase
pyruvate kinase
30

Pengaturan Pertama
Glikolisis:
Heksokinase

Mengkatalisis semua heksosa.

Terdapat hampir dalam semua sel,
kecuali hepar dan sel- pankreas.
Tidak dipengaruhi oleh;
puasa,
diet,
insulin,
diabetes
melitus
Dihambat oleh Glukosa-6-fosfat.
31

Pengaturan Pertama
Glikolisis:
Glukokinase

Mengkatalisis hanya glukosa saja.

Terdapat dalam sel hepar saja.
Dipengaruhi oleh;
puasa,
diet,
insulin,
diabetes
melitus
Tidak dihambat oleh Glukosa-6-fosfat.

32

Pengaturan Glikolisis
Utama:
Fosfofruktokinase

Enzim ini membatasi / mengatur

kecepatan jalur glikolitik.
Diaktifkan
oleh peningkatan AMP
dalam sitosol.
AMP meningkat karena ATP dihidrolisis
oleh reaksi yang memerlukan energi.

33

Pengaturan oleh Piruvat Kinase

Piruvat Kinase diatur didalam hepar.

Enzim ini mengubah kelebihan glukosa
menjadi
piruvat,
selanjutnya
dimetabolisme menjadi acetyl-CoA agar
bisa disimpan sebagai fatty acids untuk
cadangan energi jangka panjang.
34

Hasil Bersih Glikolisis Aerob

Glukosa + 2NAD+ + 2Pi + 2 ADP

2Piruvat + 2NADH + 4H+ + 2ATP +

2H2O
35

Hasil Bersih Glikolisis

Anaerob
Glukosa + 2 ADP + 2 Pi

2 lactate + 2 ATP
36

HMP-Shunt

Disebut juga jalur pentosa fosfat /

heksosa monofosfat.
Jalur ini menghasilkan NADPH dan
ribosa di luar mitokondria.
NADPH diperlukan untuk biosintesis;
asam lemak,kolesterol, dan steroid lain.
Ribosa untuk biosintesis asam nukleat.

37

Kepentingan lain

HMP-shunt berlangsung dalam jaringan;

hepar, lemak, korteks adrenal, tiroid,
eritrosit, kelenjar mammae sedang
laktasi.
NADPH juga penting dalam;
detoksifikasi obat oleh
monooksigenase,
reduksi glutation.
38

HMP-shunt terdiri dari fase:

1.

Oksidatif (irreversible);
glukosa 6-fosfat ---> ribulosa 5fosfat

2. Non-oksidatif (reversible);
ribulosa 5-fosfat ---> ribosa 5-fosfat
39

Jalur HMP-Shunt

40

HMP-shunt terdiri dari fase:

1.

Oksidatif (irreversible);
glukosa 6-fosfat ---> ribulosa 5fosfat

2. Non-oksidatif (reversible);
ribulosa 5-fosfat ---> ribosa 5-fosfat
41

Jalur HMP-Shunt

42

Hasil Bersih
3 glukosa 6-fosfat + 6 NADP+

3 CO2 + 6 NADPH + 6H+ +

2 fruktosa 6-fosfat + gliseraldehida
3-fosfat
43

Defisiensi glukosa 6-fosfat

dehidrogenase (G6PD)

Eritrosit
matang
sudah
tidak
mengandung mitokondria,
Sehingga sangat tergantung pada G6PD.
NADPH diperlukan untuk mereduksi;
glutation teroksidasi --> glutation
tereduksi
(GSH)
-->
(GS-SG)
GSH penting untuk meredam H2O2.
44

Defisiensi glukosa 6-fosfat

dehidrogenase (G6PD)

Hidrogen
peroksida
(H 2O2)
menyebabkan Hb ---> metHb, karena
Fe2+ ---> Fe3+.
Akibatnya terbentuk badan-Heinz yang
akan menimbulkan anemia hemolitik.
Penyakit ini makin memburuk bila
penderita memakan obat malaria
primaguin atau kacang fava.
45

Composition of Blood:
White Blood Cells

White blood cells

AKA: Leukocytes or WBCs

Largest sized blood cells

Lowest numbers in the blood

(4,500 11,000 per microliter)

Formed in the bone marrow

and some in lymph glands

Primary cells of the immune system

Fights disease and foreign invaders

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Composition of Blood:
White Blood Cells

White blood cells

Contain nuclei with DNA,
the shape depends on type of cell

Certain WBCs produce antibodies

Life span is from 24 hours to several years

Size is 8-20 micrometers in diameter

There are five different types of WBCs

Neutrophils
Eosinophils
Basophils
Lymphocytes
Monocytes

1.
2.
3.
4.
5.

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Composition of Blood:
Platelets

Platelets
AKA: Thrombocytes or PLTs

Formed in the bone marrow

Fragments from the cytoplasm of megakaryocytes

Smallest of the blood cells

1-4 micrometers in diameter

Shape can be round, oval, or appear spiky

Life span of around 8-12 days

Composition of Blood:
Platelets

Platelets
Involved in the clotting process

Seal wounds and prevent blood loss

Help repair damaged vessels

150,000 400,000 per microliter of blood

Platelets stain bluish with reddish or purple granules

Metabolism of
leukocyte

Differentiation of the bone marrow stem

cells
myeloid
progenitor

stem cell

megakaryocyte
erythroblast

lymphoid
progenitor

erythrocyte

monocyte
platelets
neutrophil eosinophil basophil

dendritic cell

macrophage
mature
lymphocytes

plasma
cell

A) Phagocytic cells:

Neutrophils most abundant

Eosinophils
Monocytes
Macrophages rise by differentiation of monocytes in
tissues

Degradation of the ingested

particle:

1) Activation of NADPH oxidase

2) Production of NO by nitric oxide synthase

3) Fusion of phagosome with lysosomes of the phagocytic cell

that contain bactericidal substances and hydrolytic enzymes
(often with acidic pHopt)

1) NADPH-oxidase

Protein complex of neutrophils, eosinophils, monocytes,

macrophages
superoxide anion
NADPH + 2 O2 NADP+ + H+ + 2 O22 O2- + 2 H+ O2 + H2O2

H2O2 can damage bacteria directly or after conversion to OH :

H2O2 + M+ OH + OH- + M2+ (M; metal)

Activation: by association of the components localized in

cytosol with cytochrome b558 in the membrane; electrons
from cytosolic NADPH are via FAD and cytochrome
transferred to oxygen

cytochrome b558
active NADPH-oxidase

plasma
membrane
fusion
with
lysosomes
phagosome

Myeloperoxidase

Present in granules of neutrophils and monocytes, but not

macrophages!

Significant portion of H2O2 (produced by dismutation of O 2- generated

by NADPH oxidase) is used by myeloperoxidase to oxidize Cl- to HClO

HClO is highly reactive, able to oxidize biomolecules; it also provides

toxic chlorine gas:
HClO + H+ + Cl- Cl2 + H2O

HClO also reacts with O2- yielding OH:

HClO + O2- O2 + OH + Cl-

Chronic granulomatous disease

Caused by a deficiency of one of the NADPH oxidase subunits

Superoxide and the other reactive oxygen species are not

produced

Severe infections that are very hard to treat e.g.:

Burkholdaria cepacea causes pneumonia

Aspergillus causes intractable pneumonia, septicaemia; can
lead to death

Treatment: antibiotics, antifungal agents

2) Nitric oxide production

Mainly by inducible nitric oxide synthase (iNOS) of macrophages which

is induced by cytokines (INF-, TNF) or bacterial lipopolysaccharide:

Arg

citrulline

NO can kill bacteria directly (e.g. by inhibition of the respiratory

chain) or indirectly: by reaction with O2-, generating peroxynitrite
ONOO- which attacks Fe-S proteins and essential SH groups,
inactivates enzymes

NADPH oxidase is effective mainly in degradation of

extracellular pathogens (Salmonella, Staphylococcus,
Streptococcus pyogenes)neutrophils
X

NO serves mainly to kill the intracellular parasites (Listeria,

Brucella, Candida albicans)macrophages

3) Granules (lysosomes) of
neutrophils

Contain bactericidal substances and hydrolases that, after

fusion with phagosome, destroy the engulfed particles:
myeloperoxidase
lysozyme cleaves glycosidic bonds in peptidoglycan of the
bacterial (primarily G+) cell walls
defensins cationic peptides (Arg) with M of 3,5-6 kDa;
r
interact with anionic lipids of bacterial membrane and make
pores in it; can also inhibit synthesis of DNA and proteins
hydrolases, e.g. elastase serine protease: can damage
bacteria and cleave virulence factors, but also cause harm
to host tissues (cleaves the proteins of extracellular matrix,
too)

Eosinophils

Main task: defence against multicellular parasites

Display all the above-mentioned mechanisms with slight
differences:
ROS production
peroxidase of eosinophils similar to myeloperoxidase, but
prefers Br- as a substrate (instead of Cl-), thus generating
HBrO (instead of HClO)
basic protein of eosinophils disrupting the parasite cell
membranes

B) Basophils and mast

cells

Activated by antigens / allergens interacting with IgE bound to the surface IgE receptors of basophils (mast cells)
Upon activation, content of their granules is released substances that are harmful to parasite and induce reactions that should lead to its
removal; however, they can also be responsible for allergic symptoms:
hydrolases
histamine
heparin

Synthesis of eicosanoids is activated; leukotrienes are potent bronchoconstrictors, stimulate chemotaxis and leukocyte activation

cytoplasmic granules

Histamine

Produced by histidine decarboxylation:

Causes vasodilation and bronchoconstriction helps to

eliminate parasites (cough, peristalsis, enhanced production of
mucus)

Atopy

IgE recognizing allergens (from pollen, food) are produced and

bind to IgE receptors of basophils (mast cells). Next exposure to
the allergen can lead to release of histamine and heparin and
synthesis of eicosanoids

Local symptoms occur: allergic rhinitis, asthma, conjunctivitis

If the allergen enters bloodstream, it can cause a massive

degranulation of basophils (mast cells) increase in vascular
permeability, decrease in blood pressure pulmonary oedema,
ischemia anaphylactic shock

Treatment: antihistamines block histamine receptors

C) Lymphocytes

Have specific receptors recognizing one particular antigen: B

cell receptors (BCR) and T cell receptors (TCR), respectively

BRC is a membrane-bound immunoglobulin, TCR is very similar

to Ig

B cells (after proliferation and differentiation into plasma cells)

secrete large amounts of antibodies (soluble immunoglobulins)

Soluble immunoglobulins

VH

VL

CH1

Fab

CL

CH2

Fc
CH3

2 heavy chains (H) interconnected

by disulfide bonds
2 light chains (L), each connected
to one of the H chains (by disulfide
bond)
H chain: 4-5 domains, 50-75 kDa
L chain: 2 domains, 25 kDa
N-terminal domains of H- and Lchains are variable (VH resp. VL),
the others are constant (CH resp.
CL), i.e. the same in one type of Ig
Variable domains of H a L chains
form the antigen-binding site

Types of immunoglobulins
(Ig)
155 kDa

(similar: IgD, IgE)

900 kDa

There are 2 isotypes of L: ,

There are 5 isotypes of H:
, , , ,
According to these isotypes of
H, 5 types of immunoglobulins
can be distinguished:
IgA (2 subtypes)
IgG (4 subtypes)
IgD
IgE
IgM
IgM can form pentamer, IgA
can form dimer or trimer

D) Platelets

No nucleus many of their metabolites come from

megakaryocytes
Form blood clots, act as vasoconstrictors
Participate in defence against infections, e.g.: they suppress the
growth of Plasmodium falciparum (infectious agent that causes
malaria)
Generate O2- and H2O2 that may synergize with proaggregatory stimuli
Contain thromboxan A synthase that catalyzes conversion of
prosta-glandin H2 to thromboxan A2:
TXA2 promotes platelet aggregation and vasoconstriction

Platelets also release two very important factors that can

influence not only platelets but also other cell types:

Platelet-Activating Factor (PAF)

Platelet-Derived Growth Factor (PDGF)

Platelet-Activating Factor
phospholipid

Mainly juxtacrine and paracrine signalling via GPCR

Promotes platelet aggregation
Induces activation of leukocytes, adhesion, chemotaxis, cytokine
production, causes vasodilation and bronchoconstriction
Mediates interplay between thrombotic and inflammatory cascades
BUT: it is also suspected of contributing to allergy, anaphylactic shock
It is produced also by endothelial cells, monocytes, granulocytes

Platelet-Derived Growth Factor

Dimeric protein, 3 isoforms

Receptors: tyrosine kinases expressed on fibroblasts, glia,
smooth muscle cells, leukocytes.
Effects:
proliferation
chemotaxis
cytoskeletal rearrangements
differentiation of certain types of cells (e.g. in CNS)
participates in wound healing, capillary formation,
embryonic and postnatal development!
BUT: probably also plays a role in pathogenesis (some tumours)

Cytokines

Proteins secreted by leukocytes and other cells (but there are

also membrane cytokines) that influence (via receptors) the cells
of the immune system

Cytokine signalling:
autocrine a cytokine influences the same cell that produces
it
paracrine a cytokine influences the nearby cells
endocrine a cytokine influences distant cells (after transport
by the bloodstream)

Types of cytokines

Interleukins e.g. IL-6: produced by macrophages, neutrophils, stimulates lymphocytes, secretion of Ig, synthesis of acute phase reactants

Chemokines induce chemotaxis

Interferons e.g. INF-: produced by lymphocytes, monocytes, and

macrophages, participates in antiviral defense (induces synthesis of
enzymes that block viral replication)

Transforming growth factors e.g. TGF-: produced by Tlymphocytes, macrophages, and platelets, displays anti-inflammatory
effects

Tumor necrosis factors e.g. TNF-: able to induce apoptosis

Leukocyte infiltration into tissues

= diapedesis (extravasation):

Taken from:
Halliwell, Gutteridge,
Oxford University Press, 1999

Leukocytes are slowed down by the interaction of their mucins with

selectines on the surface of endothelial cells (EC)
Cytokines on the surface of EC interact with the receptors of leukocytes
A strong adhesion mediated by the interaction of integrins with
molecules on the surface of EC migration of leukocytes into the tissue
directed by cytokins released by inflammatory cells or EC

Regulation

Many functions of leukocytes are regulated by monomeric GTPbinding proteins, e.g. Rac, Rho:
activation of NADHP oxidase
chemotaxis
phagocytosis
fusion of phagosome with granules

Rho and Rac are able to modulate the assembly of actin

filaments, which plays a role in the processes listed above

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Hemoglobin
Structure & Function

Objectives of the
Lecture
1- Understanding the main structural &
functional details of hemoglobin as one of the
hemoproteins.
2- Identify types & relative concentrations of
normal adult hemoglobin with reference to
HBA1c with its clinical application.
3- Recognize some of the main genetic &
biochemical aspects of methemoglobinopathies
with some implications on clinical features
(with focusing on thalassemias).
thalassemias

Hemoglobin is a globular
hemoprotein

Hemeproteins are a group of specialized proteins that contain

heme as a tightly bound prosthetic group.

Heme is a complex of protoporphyrin IX and ferrous iron

(Fe2+) .

The iron is held in the center of the heme molecule by bonds to

the four nitrogens of the porphyrin ring.

The heme Fe2+ can form two additional bonds, one on each
side of the planar porphyrin ring.
In myoglobin and hemoglobin, one of these positions is
coordinated to the side chain of a histidine residue of the globin
molecule, whereas the other position is available to bind oxygen

Globin of hemoglobin is a globular

protein with a quaternary structure

Structure of heme
Heme is a complex of protoporphyrin IX and ferrous
iron (Fe2+).
The iron is held in the center of the heme molecule
by bonds of the four nitrogens of the protoporphrin
ring.
Heme F2+ can form two additional bonds, one on each
side of
the porphyrin ring. One of these positions is
coordinated to the
Side chain of histidine residue of the globin molecule,
whereas
the other position is available to bind oxygen.
oxygen

Structure & function of

hemoglobin

Hemoglobin is found exclusively in RBCs.

Its main function is to transport oxygen from lungs to the
tissues & carbon dioxide & hydrogen protons from tissues
to lungs.
Hemoglobin A is the major hemoglobin in adults, is
composed of four polypeptide chains, 2 alpha () & 2
beta () chains, held together by noncovalent interactions
Each subunit has stretches of -helical structure & a
heme binding pocket.

Structure & function of

hemoglobin (cont.)

Structure & function of

hemoglobin (cont.)
Quaternary structure of hemoglobin:

The hemoglobin tetramer can be envisioned as being composed of two identical

dimers, ()1 and ()2, in which the numbers refer to dimers one and two.

The two polypeptide chains within each dimer are tightly held together,
primarily by hydrophobic interactions

In contrast, the two dimers are able to move with respect to each other, being
held together primarily by polar bonds.

The weaker interactions between these mobile dimers result in the two dimers
occupying different relative positions in deoxyhemoglobin as compared with
oxyhemoglobin

oxygenation & deoxygenation of

hemoglobin
(oxyhemoglobin & deoxyhemoglobin)

Deoxyhemoglobin
Taut structure

Oxyhemoglobin
Relaxed structure

Types of adult hemoglobin

% 6 3

HBA:
HBA2:
adult HB
HBF:
HBA1C :
amounts

the major hemoglobin in humans

first appears 12 weeks after birth- a minor component of normal
normally synthesized only during fetal development
has glucose residues attached to -globin chains increased
in DM

Hemoglobin A1c (HBA1c)

Some of hemoglobin A is
glycosylated
Extent of glycosylation depends on
the plasma concentration of a
. particular hexose (as glucose)
The most abundant form of
glycosylated hemoglobin is HBA1c
which has a glucose residues
attached to -globin chains in
.hemoglobin RBCs
Increased amounts of HBA1c are
found in RBCs of patients with
HbA1c could be used as a monitor for the control of the
.diabetes mellitus (DM)
blood glucose level during the last 2 months for diabetic
patients

Hemoglobinopathies
Hemoglobinopathies are members of a family of genetic
disorders caused
by:

1- Production of a structurally abnormal hemoglobin

molecule
(Qualitative hemoglobinopathies)
Or: 2- Synthesis of insufficient quantities of normal
hemoglobin
(Quantitative hemoglobinopathies)
Or: 3- both (rare).
(rare)

Thalassemias

Thalassemias are hereditary hemolytic diseases in which

an imbalance occurs in the synthesis of globin chains.

They are most common single gene disorders in

humans.

Normally,
Normally synthesis of - and - globin chains are
coordinated, so that each -globin chain has a -globin
chain partner.
This leads to the formation of 22 (HbA).
In thalassemias,
thalassemias the synthesis of either the - or globin chain is defective.

Thalassemias

(cont.)

Thalassemia can be caused by a variety of mutations, including:

1- Entire gene deletions (whole gene is absent)
absent
Or: 2- Substitutions or deletions of one or more nucleotides in
the DNA.

Each thalassemia can be classified as either:

either
1- A disorder in which no globin chains are produced
( o- or o -thalassemia)

Or: 2- Some -chains are synthesized, but at a reduced rate.

rate
( +- or +- thalassemia).

Thalassemias

(cont.)

1- -thalassemias:

Synthesis of -globin chains are decreased or absent, whereas

-globin
chain synthesis is normal.
-globin chains cannot form stable tetramers, and therefore
precipitate
causing premature death of RBCs.
RBCs
Accumulation of 2 2 (HbF), 4 (Hb Bart's) & -chain precipitate
occurs.
These factors end result in development of chronic anemia
(hemolytic).

Thalassemias

(cont.)

Some genetic aspects of thalassemia:

There are only two copies of the -globin gene in each cell (one on
each
chromosome 11).
So, individuals with -globin gene defects have either:

1- -thalassemia minor ( -thalassemia trait):

if they have only one defective -globin gene.
2- - thalassemia major (Cooley anemia):
if both genes are defective.

Thalassemias

(cont.)

Mutation in one of
-globin genes

Mutation in both
-globin genes

-thalassemia
minor

-thalassemia
major

Thalassemias

(cont.)

Some clinical aspects of

thalassemias:
1- As -globin gene is not expressed until late fetal gestation, the physical
manifestations of - thalassemias appear only after birth.
2- Individuals with - thalassemias minor, make some -chains, and
usually
require no specific treatment.

3- Infants born with - thalassemias major seem healthy at birth, but

become severely anemic during the first or second years of life.
They require regular transfusions of blood.
In these cases, bone marrow replacement therapy is recommended .

Thalassemias

(cont.)

2- -thalassemia:

Synthesis of -globin chains is decreased or absent.

Each individual's genome contains four copies of the
-globin (two
on each chromosome 16), there are several levels of
-globin chain
deficiencies

Thalassemias

(cont.)

Types:

If one of the four genes is defective

the individual is termed a silent carrier of - thalassemia as no physical manifestations
of the
disease occur.

If two -globin genes are defective,

the individual is designated as having -thalassemia trait.

If three -globin genes are defective;

the individual has hemoglobin H (HbH) disease which is a mildly to moderately hemolytic
anemia.
Synthesis of unaffected - and then - globin chains continues, resulting in the accumulation
of
tetramer in the newborn ( 4, Hb Bart's) or -tetramers ( 4, HbH).
The subunits do not show heme-heme interactions. So, they have very high oxygen
affinities. Thus,
they are essentially useless as oxygen carriers to tissues

If four -globin genes are defective,

hydrops fetalis & fetal death (death at birth), occurs as -globin chains are required for the
synthesis of HbF

Thalassemias

(cont.)

Types of -thalassemias

Assignments

Methemoglobin

Sickle cell anemia (Genetic,

Biochemical & Clinical Aspects)

BIOSINTESIS HEMOGLOBIN
(PORFIRIN)

Struktur Porfirin

Porfirin adalah senyawa siklik yg

dibentuk oleh 4 cincin pirol.
Masing-masing cincin dihubungkan oleh
4 jembatan metenil (-HC=).
Sifat
khas
porfirin
adalah
atom
nitrogennya mampu mengikat ion
logam.
Contoh;
- heme pada Hb mengikat Fe
- klorofil pada tumbuhan hijau mengikat
102
Mg

Struktur Porfirin
(C20H14N4)

103

Beberapa Hemoprotein
Protein
- Hemoglobin
darah
- Mioglobin
- Sitokrom c
elektron
- Sitokrom
P450
obatan
- Katalase
(H2O2)
- Triptofan
pirolase

Fungsi
mengangkut oksigen di dalam
menyimpan oksigen di dalam otot
keterlibatan pada rantai transpor

hidroksilasi xenobiotik/obatdegradasi hidrogen peroksida

oksidasi triptofan

104

Sintesis Heme di Mitokondria

85% sintesis heme terjadi dalam sel

pembentuk eritrosit pada sumsum
tulang
Heme disintesis dari suksinil KoA +
glisin.
Piridoksal
fosfat
diperlukan
untuk
mengaktifkan glisin.
Hasil kondensasi tsb ialah;
asam -amino--keto-adipat
Kondensasi
diatas
dikatalisis
oleh
Aminolevulinat-sintase (ALA-sintase). 105

Sintesis Heme di Mitokondria

Asam
-amino--keto-adipat
dengan
cepat mengadakan dekarboksilasi untuk
membentuk -aminolevulinat (ALA).
Reaksi ini dikatalisis oleh ALA-sintase.
ALA-sintase adalah enzim pengendali
laju reaksi biosintesis porfirin di hepar.

106

Sintesis Heme di Mitokondria

107

Sintesis Heme di Sitosol

Dua molekul ALA berkondensasi melalui

kerja enzim ALA-dehidratase.
Produk;
- 1 mol.porfobilinogen (PBG)
- 2 mol. H2O
ALA-dehidratase mengandung seng (Zn).
Enzim ini dapat diinhibisi oleh timbal
(Pb),
sebagaimana
terjadi
pada
keracunan Pb.
108

Sintesis Heme di Sitosol

109

Sintesis Heme di Sitosol

Kondensasi
4
mol.PBG
menghasilkan
tetrapirol linier, yaitu hidroksimetilbilana.
Reaksi ini dikatalisis oleh uroporfirinogen-1sintase (PBG deaminase).
Hidroksimetilbilana mengalami siklisasi
spontan membentuk uroporfirinogen I atau,
Menjadi uroporfirinogen III yang dikatalisis
oleh uroporfirinogen III kosintase.

110

111

Sintesis Heme di Sitosol

Uroporfirinogen III dikatalisis oleh

enzim uroporfirinogen dekarboksilase
menjadi koproporfirinogen III.
Pada
penderita
porfiria,
uroporfirinogen dekarboksilase juga
bisa mengubah Uroporfirinogen I jadi
koproporfirinogen I.
Koproporfirinogen
III
selanjutnya
memasuki mitokondria.
112

Sintesis Heme di Sitosol

113

Sintesis Heme di Mitokondria

Koproporfirinogen
III
memasuki mitokondria.

selanjutnya

Koproporfirinogen
oksidase
mengkatalisis dekarboksilasi senyawa
tsb menjadi protoporfirinogen III (IX).

Enzim ini hanya mampu

untuk koproporfirinogen III.

bekerja
114

Sintesis Heme di Mitokondria

115

Sintesis Heme di Mitokondria

Protoporfirinogen III akan dioksidasi

oleh
protoporfirinogen
oksidase
menjadi protoporfirin III (IX).

Terakhir, penyatuan ion Fe2+ (ferro)

pada protoporfirin III yang dikatalisis
oleh ferokelatase/heme sintase agar
menjadi heme.
116

Sintesis Heme di Mitokondria

117

Sintesis Heme di Mitokondria

118

Sintesis Heme

119

Pengaturan Sintesis Heme

Enzim regulator adalah ALA-sintase.

Heme bertindak sebagai regulator

negatif (umpan balik negatif) sintesis
enzim ALA- sintase.

Jika heme meningkat, maka sintesis

ALA-sintase akan menurun.
120

Beberapa Faktor Yang

Mempengaruhi
Sintesis Heme
1.

2.

Metabolisme obat-an di sitokrom P450 akan banyak menghabiskan heme

intrasel, akibatnya sintesis heme akan
meningkat.
Glukosa & hematin dapat mencegah
sintesis ALA-sintase.
121

Sifat Porfirin

Berbagai porfirinogen tidak berwarna.

Sedangkan semua porfirin berwarna,
karena adanya ikatan rangkap yang
menyatukan cincin pirol.
Porfirin yg terlarut dalam asam mineral
kuat atau pelarut organik disinari dgn
UV, maka akan mengeluarkan cahaya
fluorecen merah.
Sifat porfirin ini digunakan untuk
menegakkan diagnosis porfiria dengan
menggunakan spektrofotometer.
122

Porfirin pada Sel Kanker

Sel kanker tertentu mengambil lebih

banyak porfirin daripada sel normal.
Sifat fotodinamik porfirin dimanfaatkan
untuk fototerapi kanker.
Metode terapi;
- Penderita tumor diberi hematoporfirin,
- kemudian tumor tsb disinari dengan laser
argon yang akan memicu porfirin menjadi
sitotoksik.
123

Porfiria

Merupakan
gangguan
genetik
biosintesis heme.
Umumnya autosomal dominan, kecuali
porfiria eritropoitik kongenital.
Gejala;
- nyeri abdomen
- gangguan neuropsikiatri
- fotosensitifitas kulit
- bila berat = prototipe manusia srigala
124

Dasar Biokimia Porfiria

Nyeri abdomen & neuropsikiatri

mungkin
akibat
ALA
dapat
menghambat
enzim
ATPase
di
jaringan saraf atau,

ALA mungkin diambil oleh jaringan

otak sehingga melumpuhkan hantaran
impuls saraf.
125

Dasar Biokimia Porfiria

Fotosensitifitas
disebabkan
oleh
akumulasi porfirinogen yg mudah
teroksidasi di kulit.
Bila
terpajan
cahaya
tampak
(400nm), maka porfirin akan terpicu
untuk
bereaksi
dengan
oksigen
molekular membentuk radikal oksigen.
Radikal oksigen dapat merusak lisosom
& organel lain mengeluarkan enzim
pengurai yang merusak kulit.

126

Dasar Biokimia Porfiria

Mutasi DNA
Abnormalitas enzim pada sintesis heme
Akumulasi ALA & PBG atau
porfirinogen
penurunan heme dlm sel &
tubuh
Cairan tubuh
Tanda & gejala
porfirinogen
neuropsikiatrik

Akumulasi
di kulit & jaringan

Oksidasi spontan
menjadi porfirin
Fotosensitifitas

127

Terapi Porfiria

Hanya simptomatik.
Represor ALA-sintase;
- glukosa
- hematin (bentuk hidroksida dari
heme)
- -karoten untuk fotosensitifitas
- preparat tabir surya
Kontraindikasi;
- preparat anestesi
- alkohol
- griseofulvin & barbiturat
128

Tipe Porfiria
1.

Anemia sideroblastik terangkai-X

(eritropoitik)
- defisiensi ALA-sintase
- gejala: anemia
- Lab: hitung eritrosit & hemoglobin menurun

2.

Defisiensi ALA-dehidratase (hepatik)

- gejala: nyeri abdomen, neuropsikiatrik
- Lab: ALA urine positif

3.

Porfiria akut intermiten (hepatik)

- defisiensi uroporfirinogen-1-sintase
- gejala: nyeri abdomen, neuropsikiatrik
- Lab: PBG & uroporfirin urine positif

129

Tipe Porfiria
4. Eritropoitik kongenital (eritropoitik)
- defisiensi uroporfirinogen-III-sintase
- gejala: tanpa fotosensitifitas
- Lab: uroporfirin urine positif & PBG urine negatif
5. Porfiria kutanea tarda (hepatik)
- defisiensi uroporfirinogen dekarboksilase
- gejala: fotosensitifitas
- Lab: uroporfirin urine positif & PBG urine negatif
6. Koproporfiria herediter (hepatik)
- defisiensi koproporfirinogen oksidase
- gejala: fotosensitifitas, nyeri abdomen,
neuropsik
- Lab: PBG & uroporfirin urine positif
protoporfirin feses positif

130

Tipe Porfiria
7. Porfiria variegata (hepatik)
- defisiensi protoporfirinogen oksidase
- gejala: fotosensitifitas, nyeri abdomen,
neuropsikiatrik
- Lab: PBG urine positif
protoporfirin feses positif
8. Protoporfiria (eritropoitik)
- defisiensi ferrokelatase
- gejala: fotosensitifitas
- Lab: protoporfirin feses positif
protoporfirin sel darah merah positif
131

Kepustakaan

Marks, DB., Marks, AD., Smith CM. 1996. Basic medical

biochemistry: a clinical approach. Dalam: B.U. Pendit,
penerjemah.
Biokimia Kedokteran Dasar: Sebuah
Pendekatan Klinis. Eds. J. Suyono., V. Sadikin., L.I.
Mandera. Jakarta: EGC, 2000.
Murray, RK. 2003. Porfirin dan pigmen empedu. Dalam:
Andry Hartono, penerjemah. Harpers Biochemistry. 25th
ed. Eds. R.K. Murray, D.K. Granner, P.A. Mayes, V.W.
Rodwell. McGraw-Hill Companies, New York: 342 - 9.
Cohen,
RJ.
Carbohydrate
metabolism.PPT.
www.med.ufl.edu/biochem/rcohen/rcohen.html. 20
07
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Hardjasasmita, P. Ikhtisar: biokimia dasar B. Balai
Penerbit FKUI. Jakarta. 1993.
Schumm, DE. Essentials of biochemistry. Dalam: Moch.
Sadikin, penerjemah. Intisari Biokimia. Jakarta: Bina
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