Professional Documents
Culture Documents
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
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
3
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Heme
Synthesis
Pathway
Endalamaw T
8/25/2014
OOC
CH2
CH2
S-CoA +
succinyl-CoA
-Aminolevulinic
Acid Synthase
OOC
CH2
NH3+
glycine
H+
CoA-SH
CO2
O
OOC
Endalamaw T
CH2
CH2
CH2
NH3+
-aminolevulinate (ALA)
8/25/2014
Endalamaw T
8/25/2014
COO
CH2
CH2
CH2
CH2
+
O
PBG Synthase
2 H2O
O
CH2
CH2
NH3+
NH3+
2 -aminolevulinate
(ALA)
Endalamaw T
H2C
COO
COO
CH2
CH2
CH2
CH
NH3+
N
H
porphobilinogen
(PBG)
8/25/2014
N
H
pyrrole
COO
CH2
CH2
CH2
H2 C
NH3+
N
H
Porphobilinogen (PBG)
Endalamaw T
8/25/2014
group.
COO
CH2
Enz
11
Endalamaw T
COO
CH2
COO
CH2
CH2
N
H
dipyrromethane
CH2
CH2
N
H
PBG Deaminase
PDB
1PDA
8/25/2014
12
Endalamaw T
8/25/2014
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 -
13
Endalamaw T
CH2 CH2
CH2
COO -COO -
CH2
COO -
8/25/2014
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
14
CH 2 CH 2
CH 2
COO -COO -
CH 2
Endalamaw T
COO -
Uroporphyrinogen III
Synthase
CH 2
CH 2
CH 2
CH 2
COO -
COO -
8/25/2014
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.
15
Endalamaw T
8/25/2014
Uroporphyrinogen III
Synthase is in a cleft
Uroporphyrinogen III
PDB 1JR2
Synthase
16
8/25/2014
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
17
Endalamaw T
8/25/2014
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
18
protoporphyrin IX
uroporphyrinogen III
COO
HN
H3C
CH3
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
COO-
COO
Endalamaw T
COO
COO
8/25/2014
6. Ferrocheletase
CH2
protoporphyrin IX
CH2
CH
CH3
CH
CH CH2
H3C
NH
Fe
19
CH2
CH2
CH2
CH2
Endalamaw T
CH CH2
COO
2H
Fe
N
H3C
Ferrochelatase
N
CH3
CH3
H3C
HN
H3C
COO
++
heme
CH3
CH2
CH2
CH2
CH2
COO
COO
8/25/2014
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.
20
Endalamaw T
8/25/2014
21
Endalamaw T
8/25/2014
22
Endalamaw T
8/25/2014
23
Endalamaw T
8/25/2014
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
24
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
26
Endalamaw T
8/25/2014
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)
Endalamaw T
8/25/2014
28
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
31
8/25/2014
32
8/25/2014
Endalamaw T
8/25/2014
34
Endalamaw T
8/25/2014
UROD activity
35
Endalamaw T
8/25/2014
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.
36
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
38
Endalamaw T
8/25/2014
Bilirubin
diglucuronide is excreted in the bile.
Endalamaw T
8/25/2014
40
Endalamaw T
8/25/2014
Bilirubin Diglucuronide
41
Endalamaw T
8/25/2014
42
Endalamaw T
8/25/2014
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.
43
Endalamaw T
8/25/2014
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
44
Endalamaw T
8/25/2014
T
Endalamaw
Conjugated
bilirubin returns to the blood.
8/25/2014
46
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
48
Endalamaw T
8/25/2014
49
Endalamaw T
8/25/2014
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.
50
Endalamaw T
8/25/2014
51
Endalamaw T
8/25/2014
52
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
54
Endalamaw T
8/25/2014
55
Endalamaw T
8/25/2014
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.
56
Endalamaw T
8/25/2014
57
Endalamaw T
8/25/2014
58
Endalamaw T
8/25/2014
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
59
Endalamaw T
8/25/2014
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.
60
Endalamaw T
8/25/2014
61
Endalamaw T
8/25/2014
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-.
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
64
Endalamaw T
8/25/2014
65
Endalamaw T
8/25/2014
66
Endalamaw T
8/25/2014
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.
Endalamaw
67
8/25/2014
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.
68
Endalamaw T
8/25/2014
69
Endalamaw T
8/25/2014
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
70
very
low
Endalamaw
T P50 (2.75 mmHg).
8/25/2014
71
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
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.
74
Endalamaw T
8/25/2014
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
75
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
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
77
Endalamaw T
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
8/25/2014
78
8/25/2014
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
Endalamaw T
8/25/2014
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
80
Endalamaw T
8/25/2014
81
Endalamaw T
8/25/2014
Kidney
loss
to
the
urine
in
renal
disease
Endalamaw
T
82
8/25/2014
83
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
85
8/25/2014
86
Endalamaw T
8/25/2014
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.
87
Endalamaw T
8/25/2014
Manage
neonatal septicemia and meningitis.
Endalamaw T
8/25/2014
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.
89
Endalamaw T
8/25/2014
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.
90
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
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.
93
Endalamaw T
8/25/2014
Endalamaw T
8/25/2014
Decreased levels:
Observed in acute phase reaction, pancreatitis and
pancreatic carcinoma.
95
Endalamaw T
8/25/2014
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.
96
Endalamaw T
8/25/2014
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.
97
Endalamaw T
8/25/2014
98
8/25/2014
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
levels
Endalamaw T
8/25/2014
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
100
pits
which
form endosomes .
Endalamaw
T
8/25/2014
101
Endalamaw T
8/25/2014
Thanks
102
Endalamaw T
8/25/2014