Topic 12. Porphirin Metabolism.

Rabhi Ranim 411B

Pathobiochemistry of jaundices. The theoretical questions:
1. Structure of hemoglobin. Human variants of hemoglobin.

The structure of hemoglobin is a globular protein composed of two alpha subunits and two beta
subunits, each containing a heme group. The alpha subunits are identical, as are the beta
subunits, but the two types of subunits differ slightly in their amino acid sequence. The alpha
and beta subunits are held together by noncovalent interactions such as hydrogen bonds, salt
bridges, and hydrophobic interactions.
There are several human variants of hemoglobin, each with slightly different amino acid
sequences that can affect the function of the protein. One of the most well-known variants is
sickle cell hemoglobin, which is caused by a single amino acid substitution in the beta subunit.
This substitution changes a glutamic acid residue to a valine residue, which can cause the
hemoglobin molecules to form long, rigid fibers that distort the shape of red blood cells and
cause them to become sickle-shaped. Sickle cell hemoglobin can lead to a variety of health
problems, including anemia, pain, and organ damage.
Another variant of hemoglobin is hemoglobin C, which is caused by a different amino acid
substitution in the beta subunit. Hemoglobin C can also cause the red blood cells to take on a
slightly abnormal shape, but it usually does not cause as severe of health problems as sickle cell
hemoglobin.
There are also several other less common variants of hemoglobin, such as hemoglobin E and
hemoglobin D, that can cause mild to moderate health problems depending on the specific amino
acid substitutions involved.

2. Biosynthesis of hemoglobin. Synthesis of ALA and further schematically till the formation of
Heme.

The biosynthesis of hemoglobin begins with the synthesis of a precursor molecule called delta-
aminolevulinic acid (ALA), which takes place in the mitochondria of immature red blood cells
(erythroblasts). The biosynthetic pathway for ALA is known as the C5 pathway, and it involves
several enzymatic reactions.
1. The first step of the pathway involves the condensation of glycine and succinyl-CoA,
catalyzed by the enzyme delta-aminolevulinic acid synthase (ALAS). This reaction
produces ALA.
2. ALA is then transported out of the mitochondria and into the cytoplasm, where it
undergoes a series of reactions to form porphobilinogen (PBG). This involves the actions
 2.

of several enzymes, including ALA dehydratase, porphobilinogen deaminase, and

uroporphyrinogen III synthase.
3. The four molecules of porphobilinogen are then combined to form a linear tetrapyrrole
molecule called uroporphyrinogen III. This reaction is catalyzed by uroporphyrinogen III
synthase.
4. Uroporphyrinogen III undergoes several modifications to form coproporphyrinogen III,
which is then transported back into the mitochondria.
5. Finally, coproporphyrinogen III is converted to protoporphyrin IX through a series of
reactions that involve the insertion of iron into the porphyrin ring. This reaction is
catalyzed by ferrochelatase, and the resulting protoporphyrin IX molecule is the precursor
to heme.
6. Protoporphyrin IX is then transported out of the mitochondria and into the cytoplasm,
where it combines with globin chains to form hemoglobin.
Overall, the biosynthesis of hemoglobin is a complex process that involves several enzymatic
reactions and multiple subcellular compartments. Any disruptions or mutations in this process
can result in various hemoglobinopathies, such as sickle cell disease and thalassemia.

3. Hormonal and metabolic regulation of heme synthesis. Participation of vitamins in synthesis of

heme.

Heme synthesis is regulated by both hormonal and metabolic factors. One of the key hormones
involved in regulating heme synthesis is erythropoietin (EPO), which is produced by the kidneys
in response to hypoxia (low oxygen levels). EPO stimulates the proliferation and differentiation of
erythroid progenitor cells, leading to an increase in the production of hemoglobin and heme.
Additionally, certain metabolic intermediates and enzymes are involved in the regulation of heme
synthesis.
The rate-limiting step of heme synthesis is the formation of delta-aminolevulinic acid (ALA) from
glycine and succinyl-CoA, which is catalyzed by the enzyme ALA synthase. The activity of ALA
synthase is regulated by a variety of factors, including heme itself, which can act as a negative
feedback regulator of ALA synthase activity. When heme levels are low, ALA synthase activity is
upregulated to increase heme production.
Several vitamins are also involved in the synthesis of heme. Vitamin B6 (pyridoxine) is a cofactor
for ALA synthase, and deficiency in this vitamin can lead to impaired heme synthesis. Vitamin
B12 (cobalamin) and folate (vitamin B9) are also important for heme synthesis, as they are
involved in the production of red blood cells. Vitamin C (ascorbic acid) is required for the proper
absorption of iron, which is necessary for the formation of heme.
Overall, the synthesis of heme is a complex process that is regulated by a variety of hormonal
and metabolic factors, as well as essential vitamins. Disruptions in this process can lead to
various disorders, such as porphyrias and anemia.

4. Hereditaries violations of porphyrins metabolism : erythropoetic porphyriya - Gunter’s

disease, hepatic porphyria.

Porphyrias are a group of genetic disorders that result from defects in the biosynthesis of heme,
which can lead to the accumulation of porphyrins and their precursors. The porphyrias can be
classified into two broad categories based on where the accumulation of porphyrins occurs:
erythropoietic porphyrias, which affect heme synthesis in the bone marrow and result in the
accumulation of porphyrins in red blood cells, and hepatic porphyrias, which affect heme
synthesis in the liver and result in the accumulation of porphyrins in the liver.
One example of an erythropoietic porphyria is congenital erythropoietic porphyria (CEP), also
known as Gunther's disease. This is a rare genetic disorder that is caused by mutations in the
uroporphyrinogen III synthase (UROS) gene, which is involved in the biosynthesis of heme. As a
result of the UROS deficiency, there is a buildup of porphyrins and their precursors in the bone
marrow, which can cause severe anemia, skin lesions, and other symptoms.
Hepatic porphyrias, on the other hand, are characterized by the accumulation of porphyrins in
the liver. One example of a hepatic porphyria is acute intermittent porphyria (AIP), which is
caused by a deficiency in the enzyme porphobilinogen deaminase (PBGD). This deficiency leads
to the accumulation of porphyrins in the liver, which can cause acute attacks of abdominal pain,
nausea, vomiting, and other symptoms.
Other types of hepatic porphyrias include porphyria cutanea tarda (PCT), hereditary
coproporphyria (HCP), and variegate porphyria (VP), which are caused by deficiencies in
different enzymes involved in the biosynthesis of heme in the liver. These disorders can cause a
range of symptoms, including skin lesions, liver damage, and neurological problems.

5. Catabolism of hemoglobin, formation of bilirubin and bilirubin-albumin complex.

Hemoglobin is broken down into heme and globin chains through a process known as
catabolism. Heme is then further broken down into biliverdin, which is then converted into
bilirubin by the enzyme biliverdin reductase. The bilirubin produced is transported by albumin to
the liver for further processing and excretion.
The catabolism of hemoglobin primarily occurs in macrophages, which are found in the spleen,
liver, and bone marrow. As erythrocytes (red blood cells) age and become damaged, they are
phagocytosed by macrophages, which release heme and globin chains from the hemoglobin
molecule.
The heme molecule is then processed by the enzyme heme oxygenase, which cleaves the heme
molecule into biliverdin, iron, and carbon monoxide. Biliverdin is then reduced to bilirubin by
biliverdin reductase.
Bilirubin is a yellow-orange pigment that is insoluble in water and therefore cannot be excreted
directly by the kidneys. Instead, it is transported by albumin to the liver, where it is conjugated
with glucuronic acid by the enzyme UDP-glucuronosyltransferase to form bilirubin diglucuronide,
which is water-soluble and can be excreted in the bile.
The bilirubin-albumin complex is an important step in this process, as it allows for the transport
of bilirubin from the site of production (the macrophages) to the liver, where it can be further
processed and excreted. Without albumin, bilirubin would be insoluble in blood and would
accumulate in tissues, leading to a condition called jaundice.

6. Formation of bilirubin-diglucuronide in the liver.

Bilirubin is formed in the catabolism of heme, primarily in macrophages, and is transported to the
liver bound to albumin. Once it reaches the liver, bilirubin undergoes a series of reactions that
ultimately result in its conjugation with glucuronic acid to form bilirubin-diglucuronide, a water-
soluble and excretable form of bilirubin.
The process of bilirubin conjugation in the liver is catalyzed by the enzyme UDP-
glucuronosyltransferase (UGT), which transfers a glucuronic acid molecule from uridine
diphosphate glucuronic acid (UDPGA) to bilirubin, forming bilirubin-monoglucuronide. This
reaction is reversible, and the bilirubin-monoglucuronide can be further conjugated with another
molecule of glucuronic acid to form bilirubin-diglucuronide.
Bilirubin-diglucuronide is then transported into the bile canaliculi, small channels between
hepatocytes (liver cells), and ultimately into the bile ducts for excretion into the small intestine.
In the intestine, bilirubin-diglucuronide is converted by bacterial enzymes to urobilinogen, which
can be further oxidized to form stercobilin, the brown pigment that gives feces their
characteristic color.
The formation of bilirubin-diglucuronide in the liver is a critical step in the excretion of bilirubin
from the body. Disruptions in this process can lead to a buildup of bilirubin in the blood, resulting
in jaundice, a yellowing of the skin and eyes. Various factors can interfere with bilirubin
conjugation, including liver disease, genetic disorders, and certain medications. Treatment for
jaundice depends on the underlying cause and may include addressing the underlying condition,
phototherapy, or in severe cases, exchange transfusion.

7. Transformation of bilirubin-diglucuronide in colon.

Bilirubin-diglucuronide, the water-soluble form of bilirubin that is excreted from the liver into the
bile, is further metabolized in the colon by the action of bacteria. In the colon, bilirubin-
diglucuronide is broken down into urobilinogen, which is then converted into several different
metabolites, including stercobilinogen and mesobilirubinogen.
Stercobilinogen is oxidized to stercobilin, which is a brown pigment that gives feces their
characteristic color. Mesobilirubinogen is also oxidized to form mesobilirubin, which is
responsible for the brown color of urine.
The breakdown of bilirubin-diglucuronide in the colon is a natural part of the body's process for
eliminating waste products. However, disruptions in this process can occur due to certain
medical conditions or medications. For example, if there is a blockage in the bile ducts, bilirubin
may not be able to reach the colon, leading to pale or clay-colored stools. Similarly, if there is a
disruption in the gut microbiome, such as through the use of antibiotics, the breakdown of
bilirubin in the colon may be altered, leading to changes in stool color or odor.

8. Pathobiochemistry of jaundice.
a. hemolytic(pre-hepatic) jaundice.

Jaundice is a condition characterized by a yellowing of the skin and eyes, resulting from the
accumulation of bilirubin in the body. There are several types of jaundice, each with its own
pathobiochemistry.
Hemolytic or pre-hepatic jaundice is caused by the increased breakdown of red blood cells
(hemolysis), resulting in an excessive amount of bilirubin being produced. This type of jaundice
can occur due to a variety of factors, including genetic disorders, autoimmune conditions,
infections, and medications.
In hemolytic jaundice, the breakdown of red blood cells leads to the release of hemoglobin,
which is subsequently broken down into heme and globin. The heme molecule is converted into
biliverdin and then into bilirubin by the enzyme heme oxygenase. This bilirubin is then released
into the bloodstream, where it binds to albumin and is transported to the liver for conjugation
with glucuronic acid to form bilirubin-diglucuronide, a water-soluble form that can be excreted in
the bile.
However, in hemolytic jaundice, the liver may not be able to keep up with the excessive amount
of bilirubin being produced, leading to a buildup of unconjugated bilirubin in the bloodstream.
This results in the yellowing of the skin and eyes characteristic of jaundice.
Other biochemical markers that may be present in hemolytic jaundice include elevated levels of
lactate dehydrogenase (LDH), which is released from the damaged red blood cells, and elevated
levels of reticulocytes, which are immature red blood cells that are produced in response to the
increased demand for new red blood cells.
Treatment for hemolytic jaundice depends on the underlying cause and may include addressing
the underlying condition, blood transfusions to replace the lost red blood cells, and medications
to help reduce the amount of bilirubin being produced

b. hepatocellular(hepatic) jaundice.

Hepatocellular or hepatic jaundice is caused by liver dysfunction that impairs the ability of the
liver to properly process and excrete bilirubin. This can occur due to a variety of factors,
including viral hepatitis, alcoholic liver disease, drug-induced liver injury, and genetic disorders
such as Gilbert's syndrome.
In hepatocellular jaundice, the liver is unable to conjugate bilirubin properly, leading to a buildup
of unconjugated bilirubin in the bloodstream. This results in the yellowing of the skin and eyes
characteristic of jaundice. In addition to elevated levels of unconjugated bilirubin, other
biochemical markers that may be present in hepatocellular jaundice include elevated levels of
alanine transaminase (ALT) and aspartate transaminase (AST), which are liver enzymes that are
released into the bloodstream as a result of liver damage.
Treatment for hepatocellular jaundice depends on the underlying cause and may include
addressing the underlying liver disease, such as antiviral medications for viral hepatitis or
abstinence from alcohol in the case of alcoholic liver disease. In severe cases, liver transplant
may be necessary to replace the damaged liver tissue with a healthy donor liver.

c. cholestatic(post-hepatic) jaundice.

Cholestatic or post-hepatic jaundice is caused by obstruction of the bile ducts, which prevents
bilirubin and other substances from being excreted properly from the liver into the intestine. This
can occur due to a variety of factors, including gallstones, tumors, and inflammation of the bile
ducts.
In cholestatic jaundice, the buildup of bilirubin in the liver and bloodstream is predominantly of
the conjugated form, which is unable to pass through the obstructed bile ducts and is
subsequently reabsorbed into the bloodstream. This results in the yellowing of the skin and eyes
characteristic of jaundice, as well as other symptoms such as itching, pale stools, and dark urine.
Other biochemical markers that may be present in cholestatic jaundice include elevated levels of
alkaline phosphatase (ALP), which is a liver enzyme that is released into the bloodstream as a
result of bile duct obstruction, and elevated levels of gamma-glutamyl transferase (GGT), which
is another liver enzyme that is involved in the metabolism of bile.
Treatment for cholestatic jaundice depends on the underlying cause and may include surgery to
remove gallstones or tumors, medications to dissolve gallstones, or procedures to open or widen
the obstructed bile ducts. In some cases, liver transplant may be necessary if the damage to the
liver is severe or irreversible.