HEME DEGRADATION

• HEMOGLOBIN
• Hemoglobin (Hb) is the red blood pigment, exclusively found in erythrocytes.
• The normal concentration of Hb in blood in males is 14–16 g/dl, and in females 13–15 g/dl.
• Hemoglobin performs two important biological functions concerned with respiration
• Delivery of O2 from the lungs to the tissues.
• Transport of CO2 and protons from tissues to lungs for excretion.
• Structure of hemoglobin
• Hemoglobin (mol. wt. 64,450) is a conjugated protein, containing globin—the apoprotein part—and the
heme—the non-protein part (prosthetic group).
• DEGRADATION OF HEME TO BILE PIGMENTS
• Erythrocytes have a life span of 120 days. At the end of this period, they are removed from the circulation.
Erythrocytes are taken up and degraded by the macrophages of the reticuloendothelial (RE) system in the
spleen and liver.
• The hemoglobin is cleaved to the protein part globin and non-protein heme.
• About 6 g of hemoglobin per day is broken down, and resynthesized in an adult man (70 kg).
• Fate of globin :
• The globin may be reutilized as such for the formation of hemoglobin or degraded to the individual amino
acids.
• The latter undergo their own metabolism, including participation in fresh globin synthesis.
• Sources of heme :
• It is estimated that about 80% of the heme that is subjected for degradation comes from the erythrocyte and
the rest (20%) comes from immature RBC, myoglobin and cytochromes.
• Heme oxygenase :
• A complex microsomal enzyme namely heme oxygenase utilizes NADPH and O2 and cleaves the methenyl
bridges between the two pyrrole rings (A and B) to form biliverdin. Simultaneously, ferrous iron (Fe2+) is
oxidized to ferric form (Fe3+) and released.
• The products of heme oxygenase reaction are biliverdin (a green pigment), Fe3+ and carbon monoxide (CO).
Heme promotes the activity of this enzyme.
• Biliverdin reductase :
• Biliverdin’s methenyl bridges (between the pyrrole rings C and D) are reduced to methylene group to form
bilirubin (yellow pigment).
• This reaction is catalysed by an NADPH dependent soluble enzyme, biliverdin reductase.
• One gram of hemoglobin on degradation finally yields about 35 mg bilirubin. Approximately 250-350 mg of
bilirubin is daily produced in human adults. The term bile pigments is used to collectively represent bilirubin
and its derivatives.
• Transport of bilirubin to liver :
• Bilirubin is lipophilic and therefore insoluble in aqueous solution.
• Bilirubin is transported in the plasma in a bound (non-covalently) form to albumin.
• Albumin has two binding sites for bilirubin—a high affinity site and a low affinity site. Approximately 25 mg of
bilirubin can bind
• As the albumin-bilirubin complex enters the liver, bilirubin dissociates and is taken up by sinusoidal surface of
the hepatocytes by a carrier mediated active transport
• Conjugation of bilirubin
• In the liver, bilirubin is conjugated with two molecules of glucuronate supplied by UDP glucuronate.
• This reaction, catalysed by bilirubin glucuronyltransferase.
• Excretion of bilirubin into bile
• Conjugated bilirubin is excreted into the bile canaliculi.
• The transport of bilirubin diglucuronide is an active process.
• Fate of bilirubin
• Bilirubin glucuronides are hydrolysed in the intestine by specific bacterial enzymes namely alpha-glucuronidases
to liberate bilirubin.
• The latter is then converted to urobilinogen (colorless compound), a small part of which may be reabsorbed into
the circulation.
• Urobilinogen can be converted to urobilin (an yellow colour compound) in the kidney and excreted. The
characteristic colour of urine is due to urobilin.
• A major part of urobilinogen is converted by bacteria to stercobilin which is excreted along with feces.
• The characteristic brown colour of feces is due to stercobilin.

•
• JAUNDICE
• The normal serum total bilirubin concentration is in the range of 0.2 to 1.0 mg/dl.
• Of this, about 0.2-0.6 mg/dl is unconjugated while 0.2 to 0.4 mg/dl is conjugated bilirubin.
• Jaundice
• jaundice is a clinical condition characterized by yellow colour of the white of the eyes (sclerae) and skin.
• It is caused by the deposition of bilirubin due to its elevated levels in the serum.
• The term hyperbilirubinemia is often used to represent the increased concentration of serum bilirubin.
• TYPES OF JAUNDICE
• Jaundice (also known as icterus) may be more appropriately considered as a symptom rather than a disease. It is
rather difficult to classify
• jaundice, since it is frequently caused due to multiple factors. For the sake of convenience to understand,
jaundice is classified into three
• major types—
• hemolytic,
• hepatic
• obstructive.
• 1. Hemolytic jaundice :
• This condition is associated with increased hemolysis of erythrocytes (e.g. incompatible blood transfusion,
malaria, sickle-cell anemia).
• This results in the overproduction of bilirubin beyond the ability of the liver to conjugate and excrete the same.
• In hemolytic jaundice, more bilirubin is excreted into the bile leading to the increased formation of urobilinogen
and stercobilinogen.
• Hemolytic jaundice is characterized by
• Elevation in the serum unconjugated bilirubin.
• Increased excretion of urobilinogen in urine.
• Dark brown colour of feces due to high content of stercobilinogen.
• 2. Hepatic (hepatocellular) jaundice :
• This type of jaundice is caused by dysfunction of the liver due to damage to the parenchymal cells.
• This may be attributed to viral infection (viral hepatitis), poisons and toxins (chloroform, carbon tetrachloride,
phosphorus etc.) cirrhosis of liver, cardiac failure etc.
• Among these, viral hepatitis is the most common.
• Damage to the liver adversely affects the bilirubin uptake and its conjugation by liver cells.
• Hepatic jaundice is characterized by
• Increased levels of conjugated and unconjugated bilirubin in the serum.
• Dark coloured urine due to the excessive excretion of bilirubin and urobilinogen.
• Increased activities of alanine transaminase (SGPT) and aspartate transaminase (SGOT) released into circulation
due to damage to hepatocytes.
• The patients pass pale, clay coloured stools due to the absence of stercobilinogen.
• The affected individuals experience nausea and anorexia (loss of appetite).
• 3. Obstructive (regurgitation) jaundice :
• This is due to an obstruction in the bile duct that prevents the passage of bile into the intestine.
• The obstruction may be caused by gall stones, tumors etc.
• Due to the blockage in bile duct, the conjugated bilirubin from the liver enters the circulation.
• Obstructive jaundice is characterized by
• Increased concentration of conjugated bilirubin in serum.
• Serum alkaline phosphatase is elevated as it is released from the cells of the damaged bile duct.
• Dark coloured urine due to elevated excretion of bilirubin and clay coloured feces due to absence of
stercobilinogen.
• Feces contain excess fat indicating impairment in fat digestion and absorption in the absence of bile (specifically
bile salts).
• The patients experience nausea and gastrointestinal pain.
• TEST FOR JAUNDICE
• Vanden bergh reaction
• This is a specific reaction to identify the increase in serum bilirubin.
• Normal serum give negative van den bergh reaction
• DIRECT AND INDIRECT REACTIONS
• Bilirubin as such is insoluble in water while the conjugated bilirubin is soluble.
• Van den bergh reagent reacts with conjugated bilirubin and gives a purple colour immediately.
• This is referred to as direct positive van den bergh reaction.
• Addition of methanol dissolves the unconjugated bilirubin, which then gives the van den bergh reaction positive
and this is referred to as indirect positive.
• If the serum contains both unconjugated and conjugated bilirubin in high concentration the purple color is
produced immediately(direct-p) which is further intensified by the addition of alcohol(indirect-p).
• This types of reactions known as biphasic.
• Vanden bergh reaction and jaundice.
• This reaction is highly useful in understanding the nature of jaundice.
• This is due to the fact that the type of jaundice is characterised by increased serum concentration of
unconjugated bilirubin(hemolytic), conjugated(obstructive) or both of them.
• Therefore the response of van den Bergh reaction can differentiate the jaundice as follows
• Indirect positive-hemolytic jaundice
• Direct positive-obstructive jaundice
• Biphasic-hepatic jaundice
• Bilirubin in urine
• The conjugated bilirubin, being water soluble is excreted in urine.
• This is in contrast to unconjugated bilirubin which is not excreted.
• Bilirubin in urine can be detected by fouchets test or gmelins test.
• HEME DEGRADATION
• HEMOGLOBIN
• Hemoglobin (Hb) is the red blood pigment, exclusively found in erythrocytes.
• The normal concentration of Hb in blood in males is 14–16 g/dl, and in females 13–15 g/dl.
• Hemoglobin performs two important biological functions concerned with respiration
• 1. Delivery of O2 from the lungs to the tissues.
• 2. Transport of CO2 and protons from tissues to lungs for excretion.
• Structure of hemoglobin
• Hemoglobin (mol. wt. 64,450) is a conjugated protein, containing globin—the apoprotein part—and the heme—
the non-protein part (prosthetic group).
• DEGRADATION OF HEME TO BILE PIGMENTS
• Erythrocytes have a life span of 120 days. At the end of this period, they are removed from the circulation.
Erythrocytes are taken up and degraded by the macrophages of the reticuloendothelial (RE) system in the spleen
and liver.
• The hemoglobin is cleaved to the protein part globin and non-protein heme.
• About 6 g of hemoglobin per day is broken down, and resynthesized in an adult man (70 kg).
• Fate of globin :
• The globin may be reutilized as such for the formation of hemoglobin or degraded to the individual amino acids.
• The latter undergo their own metabolism, including participation in fresh globin synthesis.
• Sources of heme :
• It is estimated that about 80% of the heme that is subjected for degradation comes from the erythrocyte and the
rest (20%) comes from immature RBC, myoglobin and cytochromes.
• Heme oxygenase :
• A complex microsomal enzyme namely heme oxygenase utilizes NADPH and O2 and cleaves the methenyl
bridges between the two pyrrole rings (A and B) to form biliverdin. Simultaneously, ferrous iron (Fe2+) is oxidized
to ferric form (Fe3+) and released.
• The products of heme oxygenase reaction are biliverdin (a green pigment), Fe3+ and carbon monoxide (CO).
Heme promotes the activity of this enzyme.
• Biliverdin reductase :
• Biliverdin’s methenyl bridges (between the pyrrole rings C and D) are reduced to methylene group to form
bilirubin (yellow pigment).
• This reaction is catalysed by an NADPH dependent soluble enzyme, biliverdin reductase.
• One gram of hemoglobin on degradation finally yields about 35 mg bilirubin. Approximately 250-350 mg of
bilirubin is daily produced in human adults. The term bile pigments is used to collectively represent bilirubin and
its derivatives.
• Transport of bilirubin to liver :
• Bilirubin is lipophilic and therefore insoluble in aqueous solution.
• Bilirubin is transported in the plasma in a bound (non-covalently) form to albumin.
• Albumin has two binding sites for bilirubin—a high affinity site and a low affinity site. Approximately 25 mg of
bilirubin can bind
• As the albumin-bilirubin complex enters the liver, bilirubin dissociates and is taken up by sinusoidal surface of
the hepatocytes by a carrier mediated active transport
• Conjugation of bilirubin
• In the liver, bilirubin is conjugated with two molecules of glucuronate supplied by UDP glucuronate.
• This reaction, catalysed by bilirubin glucuronyltransferase.
• Excretion of bilirubin into bile
• Conjugated bilirubin is excreted into the bile canaliculi.
• The transport of bilirubin diglucuronide is an active process.
• Fate of bilirubin
• Bilirubin glucuronides are hydrolysed in the intestine by specific bacterial enzymes namely alpha-glucuronidases
to liberate bilirubin.
• The latter is then converted to urobilinogen (colorless compound), a small part of which may be reabsorbed into
the circulation.
• Urobilinogen can be converted to urobilin (an yellow colour compound) in the kidney and excreted. The
characteristic colour of urine is due to urobilin.
• A major part of urobilinogen is converted by bacteria to stercobilin which is excreted along with feces.
• The characteristic brown colour of feces is due to stercobilin.
• JAUNDICE
• The normal serum total bilirubin concentration is in the range of 0.2 to 1.0 mg/dl.
• Of this, about 0.2-0.6 mg/dl is unconjugated while 0.2 to 0.4 mg/dl is conjugated bilirubin.
• Jaundice
• jaundice is a clinical condition characterized by yellow colour of the white of the eyes (sclerae) and skin.
• It is caused by the deposition of bilirubin due to its elevated levels in the serum.
• The term hyperbilirubinemia is often used to represent the increased concentration of serum bilirubin.
• TYPES OF JAUNDICE
• Jaundice (also known as icterus) may be more appropriately considered as a symptom rather than a disease. It is
rather difficult to classify
• jaundice, since it is frequently caused due to multiple factors. For the sake of convenience to understand,
jaundice is classified into three
• major types—
• hemolytic,
• hepatic
• obstructive.
• 1. Hemolytic jaundice :
• This condition is associated with increased hemolysis of erythrocytes (e.g. incompatible blood transfusion,
malaria, sickle-cell anemia).
• This results in the overproduction of bilirubin beyond the ability of the liver to conjugate and excrete the same.
• In hemolytic jaundice, more bilirubin is excreted into the bile leading to the increased formation of urobilinogen
and stercobilinogen.
• Hemolytic jaundice is characterized by
• Elevation in the serum unconjugated bilirubin.
• Increased excretion of urobilinogen in urine.
• Dark brown colour of feces due to high content of stercobilinogen.
• 2. Hepatic (hepatocellular) jaundice :
• This type of jaundice is caused by dysfunction of the liver due to damage to the parenchymal cells.
• This may be attributed to viral infection (viral hepatitis), poisons and toxins (chloroform, carbon tetrachloride,
phosphorus etc.) cirrhosis of liver, cardiac failure etc.
• Among these, viral hepatitis is the most common.
• Damage to the liver adversely affects the bilirubin uptake and its conjugation by liver cells.
• Hepatic jaundice is characterized by
• Increased levels of conjugated and unconjugated bilirubin in the serum.
• Dark coloured urine due to the excessive excretion of bilirubin and urobilinogen.
• Increased activities of alanine transaminase (SGPT) and aspartate transaminase (SGOT) released into circulation
due to damage to hepatocytes.
• The patients pass pale, clay coloured stools due to the absence of stercobilinogen.
• The affected individuals experience nausea and anorexia (loss of appetite).
• 3. Obstructive (regurgitation) jaundice :
• This is due to an obstruction in the bile duct that prevents the passage of bile into the intestine.
• The obstruction may be caused by gall stones, tumors etc.
• Due to the blockage in bile duct, the conjugated bilirubin from the liver enters the circulation.
• Obstructive jaundice is characterized by
• Increased concentration of conjugated bilirubin in serum.
• Serum alkaline phosphatase is elevated as it is released from the cells of the damaged bile duct.
• Dark coloured urine due to elevated excretion of bilirubin and clay coloured feces due to absence of
stercobilinogen.
• Feces contain excess fat indicating impairment in fat digestion and absorption in the absence of bile (specifically
bile salts).
• The patients experience nausea and gastrointestinal pain.

•
• TEST FOR JAUNDICE
• Vanden bergh reaction
• This is a specific reaction to identify the increase in serum bilirubin.
• Normal serum give negative van den bergh reaction
• DIRECT AND INDIRECT REACTIONS
• Bilirubin as such is insoluble in water while the conjugated bilirubin is soluble.
• Van den bergh reagent reacts with conjugated bilirubin and gives a purple colour immediately.
• This is referred to as direct positive van den bergh reaction.
• Addition of methanol dissolves the unconjugated bilirubin, which then gives the van den bergh reaction positive
and this is referred to as indirect positive.
• If the serum contains both unconjugated and conjugated bilirubin in high concentration the purple color is
produced immediately(direct-p) which is further intensified by the addition of alcohol(indirect-p).
• This types of reactions known as biphasic.
• Vanden bergh reaction and jaundice.
• This reaction is highly useful in understanding the nature of jaundice.
• This is due to the fact that the type of jaundice is characterised by increased serum concentration of
unconjugated bilirubin(hemolytic), conjugated(obstructive) or both of them.
• Therefore the response of van den Bergh reaction can differentiate the jaundice as follows
• Indirect positive-hemolytic jaundice
• Direct positive-obstructive jaundice
• Biphasic-hepatic jaundice
• Bilirubin in urine
• The conjugated bilirubin, being water soluble is excreted in urine.
• This is in contrast to unconjugated bilirubin which is not excreted.
• Bilirubin in urine can be detected by fouchets test or gmelins test.