Metabolisme Eritrosit

dr Nyoman W / dr Sri Lestari Sulistyo Rini, MSc

 HAEMOGLOBIN STRUCTURE

Haemoglobin (MW 68,000) constitutes 95%

of the red blood cell’s dry weight.

65% of haemoglobin disintesis selama the

nucleated stages of RBC maturation and
35% selama the reticulocyte stage.
 Hemoglobin

• A globular protein with an embedded heme

group. (gabungan protein globin + heme)

• Synthesized in a series of steps:

– Heme : di sintesa dlm mitochondria and
cytosol of immature RBC
– Globin (protein) di sintesa o/ ribosom in the
cytosol.
– Produksi Hb terus berlanjut sampai pd early
cell development (proerythroblast to
reticulocyte) dlm sumsum tulang
 STRUCTURE OF RBC.
• Negative surface charge / ion permukaan ( - )
• Bag of fluid with dissolved substances and hemoglobin
• Membrane :
- Outer: glycoprotein coat
- Lipid bilayer (PL 55%,Cholesterol 45%)
- Inner: protein molecules cytoskeleton
– Spectrin, Actin, Ankyrin etc.
• No sub cellular particles
 Cytoskeleton of erythrocyte
Erythrocyte cytoskeleton
• provides shape, structure,
permits stretch
• 2-D lattice of proteins links to
membrane proteins:
• spectrin (a, b)
• actin
• ankyrin
• band 4.1
• membrane proteins:
• glycophorin
• band 3 protein
•Mature rbc does not synthesize
new proteins
• Gets lipids from circulating LDL
“Sickling” in Red Blood Cells
 Physiological Characteristics and
Functions of RBC
Characteristics of RBC
③ Suspension stability: it can be described by erythrocyte
sedimentation rate (ESR) which is RBC descending distance
per hour and suspension stability is inverse proportion to
ESR. (bisa mengendap dlm tes LED)
Normal value of ESR: male, 0~15 mm/h; female, 0~20 mm/h.
ESR and clinic: some diseases bring about rouleaux formation
(mainly involved in plasma component, e.g. globulin,
fibrinogen, cholesterol) and speed up ESR / kecepatan LED
 mempunyai makna klinik. Misal, me ↑ pd infeksi kronis
Normal Hb consists of globin (a tetramer of two
pairs of polypeptide chains) and four haem groups.

Each haem group contains a protoporphyrin ring plus

ferrous iron (Fe++).

The mitochondria are the main site of

protoporphyrin synthesis.

Iron is supplied from circulating transferrin and

globin chains are synthesised on ribosomes.
Finally, ferrous iron is inserted to form haem
by haem synthetase.

Atom `ferrous iron` u/ setiap molekul haem

berikatan dg bag. proximal `histidine residue`
pd cincin globin, bukan pd bag. distal histidine
residue.

Asam amino tertumpu pada loop diantara

proximal and distal histidine residues 
membentuk `haem pocket`, yg amat esensi
dlm kapasitas mengangkut O2 .
Ada 6 variasi Hb yg bisa terbentuk.

Embryonic haemoglobins: Gower1, Gower2

dan Hb Portland.

HbF : dominan u/ Hb fetal life (65-95%).

Adults have only trace amounts of HbF
(<1%).

HbA (>95%) and HbA2 (2.5-3.5%) are the

main adult haemoglobins.
Sintesa `Alpha chain` synthesis adl secara
langsung o/ 2 `α genes` (α1 n α2), pd
chromosome 16.

Beta and delta chains dihasilkan dari single

genes on chromosome 11.

The gamma chain adl langsung o/ 2 genes, G

and A, pd chromosome 11.

Alpha chains have 141 amino acids and non-

alpha chains have 146; the exact sequence of
the amino acids has been determined.
Haem mengandung 4 cincin pyrrole, dgn
atom Fe dipusatnya berikatan dgn 4 atom N

Atom Fe memiliki 2 sisi ikatan 1 berikatan

dgn globin histidine residue dan yg satunya
beriktan dg O2
Haem disintesa t u dlm mitochondria
erythroblasts, dan bbrp di dalam cytoplasm.

Awalnya secara terbatas yi adanya fusi antara

Succinil-Co A dgn Glysine o/ mediasi enzim ALA
menjadi δ-aminolaevulinic acid (ALA).

Hal ini terjadi dlm mitochondria dan bergantung

kpd ketersediaan dari vitamin B6 (pyridoxal
phosphate).

Reaksi ini distimulasi o/ erythropoietin dan

dihambat o/ haem
Step selanjutnya terjadi dalam cytoplasma.

2 Molekul ALA berfusi menjadi porphobilinogen.

Langkah sepasang enzim membentuk

uroporphyrinogen yi proses decarboxylasi
menjadi coproporphyrinogen.

Pd titik ini jalurnya kembali menuju pusat

mitokondria yi dgn terbentuknya protoporphyrin.
Globin chains: dlm haemoglobin dewasa adalah
alpha (α)
beta (β)
gamma (γ)
delta (δ).

Hb A memiliki 2 alpha dan 2 beta chains (α2β2),

Hb F  2 alpha dan 2 gamma chains (α2γ2)
Hb A2  2 alpha dan 2 delta chains (α2δ2).

α chain adalah terbanyak dari ke 3 tipe lainnya

pd Hb dewasa
 HAEMOGLOBIN FUNCTION
SDM mengangkut O2 dari paru-2 ke dlm jaringan dan
kemudian balik lagi ke paru-2 membawa CO2

Ketika molekul hemoglobin sedang diloading atau tidak

diloading dgn O2, maka molekul globinnya juga secara
individual dapat saling berhubungan satu dengan yg
lainnya
Bila O2 tdk terloadingikatan β-nya dpt dipisah dan
masuk dalam metabolite 2,3 diphospho-glycerate (2,3-
DPG) yg menghasilkan ikatan O2 yg lebih lemah.
Selama oksigenasi, 2 ikatan β dpt bergerak
bersama yg memberi spesies afinitas O2nya yg
lebih kuat.. Karna itu sekali O2 awal dpt diikat
oleh Hb, akan dpt meningkatkan afinitas ikatan
O2 dlm penentuan molekul group haem.

Hubungan antara konsentrasi O2 dlm darah

(tekanan partiil O2) dgn proporsi ikatan O2 dan
Hb (persentase ikatan O2) membentuk kurva
sigmoid yg disebut Hb oxygen-dissociation curve
.
 RBC life span and circulation
• Rate Penggantian SDM baru mendekati 3 juta ke
dalam sirkulasi perdetik
• Penggantian sebelum mengalami hemolisa
• Komponen-2 hemoglobin sendiri adalah bersiklus
– Heme stripped of iron and converted to biliverdin, then
bilirubin
• Fe yg tidak dipakai akan disimpan secara pagositose
atau dialirkan ke dalam sirkulasi darah dalam
bentuk transferin
 Life and breakage of RBC
• Life-span: 120 days, about 4 months, each RBC circulates
27 km averagely in vessels, short life-span for aged RBC
• Breakage: places are liver, spleen and lymphatic node,
and after breakage, Hb released from RBC immediately
combine with plasma α2-globulin (Hb touched protein)
which is taken in by liver for iron reuse.
• Breakage Hb,can very toxic if it get into blood, but
normally it can be metabolized into bile pigment in liver.
• Clinic relation.  icterus
 RBC breakdown
• Healthy RBC’s live about 120 days; we break down
about 174 million per minute
• RBC’s are removed from circulation by the liver and
spleen
• Broken down into heme and globin portions
• Globin is broken down into amino acids
• Iron is removed from heme and stored or recycled
• Heme is broken down into biliverdin and then into
bilirubin
Life Cycle of Red Blood Cells

Figure 18.7
Extravascular Pathway for RBC Destruction
(Liver, Bone marrow,
& Spleen)

Phagocytosis & Lysis

Hemoglobin

Globin Heme Bilirubin

Amino acids Fe2+

Amino acid pool Excreted

 Degradation of hemoglobin
• Heme is degraded to bilirubin:
• Bilirubin is conjugated to glucuronate (more soluble),excreted
• Rbc only live ~120 days
• Globin is degraded to amino acids

Figs. 7,8
 DEGRADATION OF HEME TO BILIRUBIN

 75% is derived from RBCs

 In normal adults this results in

a daily load of 250-300 mg of
P450 cytochrome
bilirubin

 Normal plasma concentrations

are less then 1 mg/dL

 Hydrophobic – transported by
albumin to the liver for further
metabolism prior to its excretion

“unconjugated” bilirubin
 Heme synthesis
• Heme synthesis begins with d-ALA:
• Decarboxylation by d-ALA synthase
• PLP is pyridoxal phosphate
• Dehydratase joins 2 d-ALA
• 4 pyrroles form porphyrinogen

Fig. 4
Figure 19.5
 Iron metabolism
• Iron metabolism:
• Transferrin carries Fe3+ to cells; stored as ferritin
• Transferrin taken up by R-mediated endocytosis
• Hemosiderin stores excess

Fig. 6
RE = reticulo-
endothelial
system
• Heme synthesis:
• Glycine, succinyl CoA form
• d-Aminolevulinic acid
• (d-ALA)
• Each heme needs 8 of each

• Final step is Fe2+

• Heme regulates:
• inhibit 1st enzyme
• repress synthesis

• Porphyria diseases from

• defective enzymes
• intermediates accumulate
• photosensitive, toxic products

•
Fig. 3
NORMAL BILIRUBIN  Uptake of bilirubin by the liver is mediated by a carrier
METABOLISM protein (receptor)

 Uptake may be competitively inhibited by other

organic anions

 On the smooth ER, bilirubin is conjugated with

glucoronic acid, xylose, or ribose

 Glucoronic acid is the major conjugate - catalyzed by

UDP glucuronyl tranferase

“Conjugated” bilirubin is water soluble and is secreted

by the hepatocytes into the biliary canaliculi

 Converted to stercobilinogen (urobilinogen)

(colorless) by bacteria in the gut

 Oxidized to stercobilin which is colored

 Excreted in feces

 Some stercobilin may be re-adsorbed by the gut and

re-excreted by either the liver or kidney
 HYPERBILIRUBINEMIA
 Increased plasma concentrations of bilirubin (> 3 mg/dL) occurs when
there is an imbalance between its production and excretion
 Recognized clinically as jaundice
 |haematinics: the normal iron cycle 2.4
Partners in Global Health Education

Iron deficiency can be identified best by assessing the appearances of the red cells on a blood film.
Iron indices in a blood sample are helpful to confirm a lack of iron. In order to interpret these
indices, it is vital to understand how the body handles iron …..
Iron is a key constituent of haemoglobin (60-70% of total body iron is
stored here) and it’s availability is essential for erythropoiesis. In iron Soluble transferrin receptors,
deficiency, there are more divisions of red cells during erythropoiesis sTfR are on the red cell surface.
than normal. As a result the red cells are smaller (microcytic) and These can be measured and are
have a reduced haemoglobin content (hypochromic). increased in iron deficiency.

In iron deficient states, bone marrow Red blood

iron is reduced. cells

Erythroid bone
marrow
(normoblasts) Some iron binds to Reticuloendothelial system;
apoferritin to form Spleen & macrophages
ferritin, a storage
compound.

Liver
3. Dying red cells
2. Iron is then attached to are recycled by
a protein, transferrin in macrophages in
the serum (plasma), the spleen and
where it is transported to Serum iron is recycled
the bone marrow for transferrin into the plasma for
haemoglobin synthesis. further use.
Fe Duodenum

1. Iron is absorbed from the small

intestine in the ferrous state START
(Fe2+; approx. 1mg/day).
 |haematinics: vitamin B12 2.4
Partners in Global Health Education

There are a number of key steps in the absorption of Vitamin B12. The two key locations are
the stomach and the terminal ilium. Dietary vitamin B12 binds with intrinsic factor (IF) in the
stomach, a transport protein produced by gastric parietal cells. The B12-IF complex then
travels through the small intestine and is absorbed by special receptors in the distal ileum.
This pathway is important when considering possible causes of Vitamin B12 deficiency.

Oesophagus
Vitamin B12 Vitamin B12 deficiency can
Causes of vitamin
ingested take up to two years to
B12 deficiency
develop as the body has
sufficient stores for this period.
1. Pernicious Stomach
anaemia
IF Intrinsic factor Pernicious anaemia: the
2. Inadequate leading cause of B12
intake deficiency. IgG autoantibodies
target gastric parietal cells
3. Poor absorption and its product IF causing an
atrophic gastritis. This results
Distal ileum in reduced secretion of
intrinsic factor and therefore
reduced B12-IF complex for
Site of B12 absorption in the distal ileum.
absorption

Click here to
return
1 Low O2 levels in blood stimulate
kidneys to produce erythropoietin.
2 Erythropoietin levels rise
in blood.
3 Erythropoietin and necessary
raw materials in blood promote
erythropoiesis in red bone marrow.

4 New erythrocytes
enter bloodstream;
function about 120 days.

Figure 17.7, step 4

5 Aged and damaged red Hemoglobin
blood cells are engulfed by
macrophages of liver, Heme Globin
spleen, and bone
marrow; the Bilirubin Iron stored Amino
hemoglobin is as ferritin, acids
broken down. hemosiderin
Iron is bound to
transferrin and released
to blood from liver as
needed for erythropoiesis.

Bilirubin is picked up from blood

by liver, secreted into intestine in
bile, metabolized to stercobilin by
bacteria, and excreted in feces.

Circulation

Food nutrients,
including amino acids, 6 Raw materials are
Fe, B12, and folic acid, made available in blood
are absorbed from
for erythrocyte synthesis.
intestine and enter
blood.

Figure 17.7, step 6

 1 Low O levels in blood stimulate
2
kidneys to produce erythropoietin.
2 Erythropoietin levels rise
in blood.
3 Erythropoietin and necessary
raw materials in blood promote
erythropoiesis in red bone marrow.

4 New erythrocytes
enter bloodstream;
5 Aged and damaged function about 120 days.
red blood cells are
engulfed by macrophages
of liver, spleen, and bone
marrow; the hemoglobin Hemoglobin
is broken down.

Heme Globin

Bilirubin Iron stored Amino

as ferritin, acids
hemosiderin
Iron is bound to
transferrin and released
to blood from liver as
needed for erythropoiesis.

Bilirubin is picked up from blood

by liver, secreted into intestine in
bile, metabolized to stercobilin by
bacteria, and excreted in feces.

Circulation

Food nutrients,
including amino acids, 6 Raw materials are
Fe, B12, and folic acid, made available in blood
are absorbed from for erythrocyte synthesis.
intestine and enter
blood.

Figure 17.7
 Key point:
Oxidant stress! O- H2O
CLICK HERE
|red cell metabolism 2.5
Partners in Global Health Education

Embden-Meyerhof 2GSH GSSG

Contents page glycolytic pathway Hexose monophosphate
shunt.
2.1. The erythrocyte: Glucose
an overview. Red cells require a
2.2. Erythropoiesis NAPD NADPH+H+ mechanism to detoxify the
2.3. The red cell waste products
structure (accumulated oxidised
2.3.1. Cell membrane substrates) of the cell. This
Glucose- 6-P 6-PG
2.3.2. DNA synthesis shunt provides this
2.4. Red cell solution. It also provides
metabolism ADP Glucose-6-phosphate
10% of glycolysis.
dehydrogenase
However this metabolic
pathway is also
ATP susceptible to pathology.
Ribulose 5-P
The glycolytic pathway
This is a sequence Fructose-6-P
of biochemical With no cell organelles
reactions in ADP
and no mitochondria the
which glucose is fully developed
metabolised to Pyruvate kinase Hexose
erythrocyte relies on this
lactate with the ATP monophosphate
aerobic pathway to gain
generation of 2 shunt
energy (ATP) for the cell.
ATP molecules
(providing energy
Lactate
for the cell).

Pyruvate kinase deficiency: In rare circumstances there are defects within the critical glycolytic enzymes. 95% of these defects are
Key point!

associated with pyruvate kinase, a key enzyme within this pathway. The result is insufficient ATP production for cell life and therefore
premature death (haemolysis).
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked disorder that is relatively common. The G6PD enzyme is a rate-
limiting step within this pathway. If deficient, haemolysis occurs when the cell is placed under oxidative stress (e.g. by oxidative drugs, fava
beans, infections) creating a potentially severe anaemia. Click OXIDATIVE STRESS for more info.
 ENERGY METABOLISM
• Less energy required
– Na + - K + pump
– Iron in Fe ++ form

• Utilize Glucose by GLUT 1

• Anaerobic respiration – Glycolysis
– Embden Meyerhof pathway

• Pentose phosphate pathway.

– Hexose monophosphate shunt
 Erythrocyte metabolism
• Erythrocyte metabolism: Only glycolysis
• ATP for Na+/K+, Ca2+
• HMP shunt makes NADPH
• G6PD is 1st enzyme
• Lifetime rbc by G6PD activity
• 2,3-BPG modulates O2 binding
• Need Fe2+ Hb bind O2;
• If ROS made Fe3+, NADH can reduce
2,3-Bisphosphoglycerate Pathway
 Glycolysis or
Embden-Meyerhof Pathway
• Generates 90- 95% of energy needed by
RBC’s
• Glucose is metabolized and menghasilkan
2 molecules of ATP (energy).
• Fungsinya mempertahankan bentuk
SDM, flexibility dan the cation pumps
 Hexose monophosphate shunt
• Metabolizes 5-10% of glucose.
• NADPH is end product
• Protects the RBC from oxidative injury.
• Most common defect is deficiency of the enzyme
glucose-6-phosphate dehydrogenase (G-6PD).
• If the pathway is deficient, intracellular oxidants can’t
be neutralized and globin denatures then precipitates.
The precipitates are referred to as Heinz bodies
Methemoglobin Reductase pathway

• Mempertahankan status besi dlm bentuk

ferrous (Fe++) state.

• In the absence of the enzyme (methemoglobin

reductase), methemoglobin accumulates  shg
tdk dapat mengangkut O2.
 Rapoport –Leubering Shunt

• Mempertahankan SDM meregulasi transport

O2 dalam situasi hipoksia atau ketidak
seimbangan asam basa.

• Mempertahan akumulasi 2,3-DPG yg sangat

esensi dlm hal O2 tension, dan pengaturan
afinitas Hb
Handling of Free (Intravascular) Hemoglobin

Purposes: 1. Scavenge iron

2. Prevent major iron losses
3. Complex free heme (very toxic)

• Haptoglobin: hemoglobin-haptoglobin complex yg

dimetabolisme dalam hepar dan limpa membentuk iron-globin
complex + bilirubin. Mencegah kehilangan Fe dalam urin.

• Hemopexin: ikatan heme bebas. Heme-hemopexin complex

diambil oleh liver dan iron-nya disimpan dlm bentuk ferritin.

• Methemalbumin: complex oksidasi dari heme + albumin.

 When the erythrocyte is destroyed within the vascular system, hemoglobin is released directly into the blood.
Normally, the free hemoglobin quickly complexes with haptoglobin, and the complex is degraded in the liver. In severe
hemolytic states, haptoglobin can become depleted, and free hemoglobin dimers are filtered by the kidney. Additionally,
with haptoglobin depletion, some hemoglobin is quickly oxidized to methemoglobin and bound to either hemopexin or
albumin for eventual degradation in the liver.