BIOCHEMISTRY & CELL BIOLOGY

SELF-INSTRUCTION MODULE #3-1

HEME METABOLISM

BY: DR. RICHARD HANN

DEPARTMENT OF BIOCHEMISTRY
2012 by Universidad Central del Caribe
All rights reserved
PURPOSE & SCOPE OF THE SELF-INSTRUCTION MODULES

Due to the limitation of faculty-student contact hours, it is not

possible to present in the classroom all of the concepts in
Biochemistry & Cell Biology that the student must learn. The
self-instruction modules (SIMs) are designed to provide,
within a student-centered electronic format, a structured
presentation of essential concepts & information that cannot
be presented in the classroom. Each SIM is a power-point
presentation that the student may access via the Blackboard
BCB course site at his or her convenience. Studying each of
the SIMs is compulsory. The material on each SIM is tested on
the subsequent examinations.
CONTENTS OF THIS MODULE:

1. HEME BIOSYNTHESIS

2. HEME DEGRADATION
 LEARNING OBJECTIVES OF THIS MODULE
Upon completion of this module the student should be able to:

1. Outline the biosynthesis of heme, including the compartmentalization

of the enzymes involved & how the pathway is regulated
2. Describe how heme biosynthesis relates to the
diseases known as the porphyrias

3. Explain the catabolism of heme & the formation,

conjugation & excretion of bilirubin
 1. HEME BIOSYNTHESIS
THE ADULT BODY MAKES 6g TO 8g
OF HEMOGLOBIN DAILY PLUS OTHER
HEME-CONTAINING PROTEINS

THIS REQUIRES THE BIOSYNTHESIS OF

OVER 300mg OF HEME DAILY

THE HEME BIOSYNTHESIS PATHWAY

IS ACTIVE IN ALL CELLS WITH
HEME
MITOCHONDRIA

MOST OF THE BODYS HEME IS SYNTHESIZED

BY ERYTHROCYTE PRECURSOR CELLS
(ERYTHROBLASTS) IN THE BONE MARROW
 HEME IS AN IRON PORPHYRIN
A PORPHYRIN IS A TETRAPYRROLE RING

HEME

A PORPHYRIN

PORPHYRINS ABSORB VISIBLE LIGHT & ARE COLORED RED TO PURPLE

THEY ARE CONJUGATED WITH ALTERNATING SINGLE & DOUBLE BONDS

THIS IS WHY HEME-CONTAINING PROTEINS LIKE

HEMOGLOBIN & MYOGLOBIN ARE RED OR PURPLE

PORPHYROS IS GREEK FOR PURPLE

 THE BIOSYNTHETIC PRECURSORS OF
PORPHYRINS ARE CALLED PORPHYRINOGENS

HEME

A PORPHYRINOGEN

PORPHYRINOGENS DO NOT ABSORB VISIBLE LIGHT & ARE COLORLESS

PORPHYRINOGENS DO NOT HAVE A CONJUGATED

DOUBLE BOND PATTERN LIKE PORPHYRINS
 THE HEME BIOSYNTHESIS PATHWAY
THE PATHWAY STARTS & ENDS
THE MIDDLE PART IS CYTOPLASMIC
IN THE MITOCHONDRION
 GLYCINE & SUCCINYLCoA ARE THE PRECURSORS OF HEME

MITOCHONDRIAL ALA SYNTHASE (ALAS) FORMS dAMINOLEVULINIC ACID (ALA)

WHICH IS TRANSPORTED OUT TO THE CYTOSOL

PLP
CO2
+
CoA Matrix

PYRIDOXAL PHOSPHATE (PLP) IS THE COENZYME OF ALAS

THE HUMAN GENOME HAS 2 ALAS GENES

ALAS1 ON CHROMOSOME 3 IS EXPRESSED IN ALL CELLS

ALAS2 ON THE X CHROMOSOME IS EXPRESSED ONLY IN BONE MARROW

ERYTHROCYTE PRECURSOR CELLS (ERYTHROBLASTS)

ALAS2 IS RESPONSIBLE FOR THE HUGE AMOUNT OF HEME

THAT MUST BE MADE FOR ERYTHROCYTE HEMOGLOBIN
 CLINICAL CORRELATION
SIDEROBLASTIC ANEMIA (SBA)

MAIN CLINICAL MANIFESTATIONS:

PALLOR, WEAKNESS, FATIGUE, DIZZINESS

HYPOCHROMIC MICROCYTIC ANEMIA

SEEN ON CBC & PERIPHERAL SMEAR

RINGED SIDEROBLASTS SEEN IN BONE MARROW

80% OF SBA PATIENTS RESPOND TO PYRIDOXINE

SBA IS CAUSED BY DEFICIENT ALA PRODUCTION RINGED SIDEROBLASTS IN A

IN ERYTHROCYTE PRECURSOR CELLS BONE MARROW SMEAR FROM A
(ERYTHROBLASTS) PATIENT WITH SBA STAINED FOR
Fe+2 WITH PRUSSIAN BLUE
DEFICIENT ALA PRODUCTION CAUSES
INADEQUATE HEME SYNTHESIS LEADING TO
HYPOCHROMIC MICROCYTIC ANEMIA

DEFICIENT ALA PRODUCTION CAUSES Fe+2 ACCUMULATION

& DEPOSITS IN PERINUCLEAR MITOCHONDRIA
 SIDEROBLASTIC ANEMIA CAN BE EITHER INHERITED OR ACQUIRED

ASBA ERYTHROBLAST
XSBA
SLC25A38
X
ALAS2

I
X
PLP
CO2
+
Pb+2 CoA Matrix

X-LINKED SBA (XSBA) IS THE MOST COMMON FORM OF SBA &

IS CAUSED BY AN INHERITED DEFICIENCY OF ALAS2

AUTOSOMAL RECESSIVE SBA (ASBA) IS CAUSED BY DEFICIENCY OF THE

MITOCHONDRIAL GLYCINE TRANSPORTER ENCODED BY GENE SLC25A38
(LOW MITOCHONDRIAL GLYCINE LEVELS DECREASE ALA PRODUCTION)

ACQUIRED SBA CAN RESULT FROM VITAMIN B6 DEFICIENCY OR FROM LEAD POISONING
B6 IS THE PRECURSOR OF PLP: THE COENZYME OF ALAS2
Pb+2 IS A SPECIFIC IRREVERSIBLE INHIBITOR OF ALAS2
 IN THE CYTOPLASM 2 ALAs CONDENSE TO FORM
A PYRROLE RING KNOWN AS PORPHOBILINOGEN (PBG)

PBG DEAMINASE THEN DEAMINATES & CONNECTS TOGETHER

4 PBGsTO FORM HYDROXYMETHYLBILANE (HMB)

2H20

4NH3

A = Acetyl (2C)
P = Propionyl (3C)
 HMB IS ASYMMETRICALLY CONDENSED Mitoch Cytosol
INTO THE TETRAPYRROLE RING
UROPORPHYRINOGEN III (UROGEN III) BY
UROPORPHYRINOGEN III COSYNTHASE

UROGEN III IS DECARBOXYLATED ON THE 4

ACETYL GROUPS BY
UROPORPHYRINOGEN OXIDASE TO GIVE
H2O
COPROPORPHYRINOGEN III (COPROGEN III)

UROPORPHYRINOGEN III COSYNTHASE

& UROPORPHYRINOGEN OXIDASE ARE
SOLUBLE CYTOPLASMIC ENZYMES

COPROGEN III IS THEN TRANSPORTED

BACK INTO THE MITOCHONDRION BY THE
TSPO TRANSLOCATOR PROTEIN 4CO2

TSPO A = Acetyl (2C)

P = Propionyl (3C)
M = Methyl (1C)
 BACK INSIDE THE MITOCHONDRION
COPROPORPHYRINOGEN III IS OXIDATIVELY
DECARBOXYLATED TO GIVE PROTOPORPHYRINOGEN IX

PROTOPORPHYRINOGEN IX IS THEN OXIDIZED

TO GIVE PROTOPORPHYRIN IX
2H+ (THE FIRST PORPHYRIN)

COPROPORPHYRINOGEN OXIDASE &

PROTOPORPHYRINOGEN OXIDASE ARE BOTH
LOCATED IN THE INTERMEMBRANE SPACE

IN THE FINAL STEP THE MITOCHONDRIAL MATRIX

ENZYME FERROCHELATASE PUTS AN ATOM OF
3H2O2
FERROUS IRON (Fe+2) INTO THE RING TO FORM HEME
3O2
2H2O +
2CO2

M = Methyl (1C)
P = Propionyl (3C)
V = Vinyl (Ethenyl) (2C) Mitochondrion
REGULATION OF HEME BIOSYNTHESIS

HEME ALLOSTERICALLY
INHIBITS ALA SYNTHASE

THIS FEEDBACK INHIBITION

PLP IS THE MAIN CONTROL OF
CO2 THE HEME BIOSYNTHESIS
+
CoA Matrix
PATHWAY

IN THE LIVER HEME ALSO REPRESSES

ALA SYNTHASE GENE TRANSCRIPTION

IN ERYTHROBLASTS
LOW CYTOSOLIC IRON CONCENTRATION ALSO
BLOCKS ALA SYNTHASE 2 mRNA TRANSLATION
 CLINICAL CORRELATION
PORPHYRIA

THE PORPHYRIAS ARE A GROUP OF INHERITED DISEASES CAUSED BY DEFECTS

IN THE ENZYMES OF THE HEME BIOSYNTHESIS PATHWAY

HIGH BLOOD LEVELS OF PORPHYRINS OR

PORPHYRIN PRECURSORS LEAD TO THEIR
EXCRETION IN THE URINE & FECES

THIS GIVES A RED OR PURPLE COLOR TO THE

URINE & IN SOME CASES TO THE FECES

L) NORMAL URINE
M) URINE FROM A PATIENT WITH
ACUTE INTERMITTENT PORPHYRIA
R) RED WINE DILUTED WITH WATER
IN SOME PORPHYRIAS HIGH PORPHYRIN LEVELS
IN THE SKIN GIVE SEVERE PHOTOSENSITIVITY
WITH RESULTING PHOTODERMATITIS

SOME PORPHYRIAS FEATURE ACUTE ATTACKS

OF ASSORTED NEUROVISCERAL SYMPTOMS

NEUROVISCERAL SYMPTOMS CAN INCLUDE

VOMITING, ABDOMINAL PAIN, CONSTIPATION,
MYASTHENIA, PERIPHERAL NEUROPATHY, PHOTODERMATITIS IN A PATIENT
CONVULSIONS, FEVER, TACHYCARDIA, WITH PORPHYRIA
HYPERTENSION & BEHAVIORAL CHANGES

MOST PORPHYRIAS SHOW AUTOSOMAL DOMINANT INHERITANCE

THESE PORPHYRIAS MANIFEST IN HETEROZYGOTES

 6 HUMAN PORPHYRIAS
DEFICIENT MAJOR CLINICAL MAIN COMPOUNDS IN
PORPHYRIA: ENZYME: MANIFESTATIONS: URINE (U) & FECES (F):

CONGENITAL UROPORPHYRINOGEN PHOTOSENSITIVITY UROPORPHYRIN I (U)

ERYTHROPOIETIC III COSYNTHASE HEMOLYTIC ANEMIA COPROPORPHYRIN I (U + F)
PORPHYRIA (CEP) (HOMOZYGOUS)
ACUTE PBG DEAMINASE NEUROVISCERAL ALA + PBG (U)
INTERMITTENT (HETEROZYGOUS) SYMPTOMS PROPHYRINS (U + F)
PORPHYRIA (AIP) ACUTE ATTACKS DURING ATTACKS

PROTOPORPHYRINOGEN NEUROVISCERAL ALA + PBG (U)

VARIEGATE SYMPTOMS UROPORPHYRIN III +
OXIDASE
PORPHYRIA PHOTOSENSITIVITY COPROPORPHYRIN III (U + F)
(HETEROZYGOUS)
(VP) ACUTE ATTACKS PROTOPORPHYRIN IX (F)

ERYTHROPOIETIC FERROCHELATASE PHOTOSENSITIVITY PROTOPORPHYRIN IX (F)

PROTOPORPHYRIA (HETEROZYGOUS) PORPHYRIN
(EPP) GALLSTONES

PORPHYRIA UROPORPHYRINOGEN PHOTOSENSITIVITY UROPORPHYRIN III (U)

CUTANEA TARDA DECARBOXYLASE LATE ONSET
(PCT) (HETEROZYGOUS)

COPROPORPHYRINOGEN NEUROVISCERAL ALA + PBG (U)

HEREDITARY
OXIDASE SYMPTOMS COPROPORPHYRIN III (U + F)
COPROPORPHYRIA
(HETEROZYGOUS) PHOTOSENSITIVITY
(HCP)
ACUTE ATTACKS
 CLINICAL CORRELATION
CONGENITAL ERYTHROPOIETIC PORPHYRIA (CEP)

MAIN CLINICAL
MANIFESTATIONS:

PHOTOSENSITIVITY

HEMOLYTIC ANEMIA WITH

SPLENOMEGALY

PORT-WINE URINE
UROPORPHYRIN I &
COPROPORPHYRIN I ARE
EXCRETED IN THE URINE
HANDS OF A PATIENT WITH
CONGENITAL ERYTHROPOIETIC
PURPLE FECES
PORPHYRIA
COPROPORPHYRIN I IS
EXCRETED IN THE FECES

INHERITANCE IS
AUTOSOMAL RECESSIVE
CEP IS CAUSED BY DEFICIENCY OF UROPORPHYRINOGEN III COSYNTHASE

IN CEP UROPORPHYRINOGEN TYPE I FORMS NONENZYMATICALLY

FROM HMB BECAUSE THE COSYNTHASE IS DEFICIENT
COPROPORPHYRINOGEN I FORMS ENZYMATICALLY FROM UROGEN I

UNCOLORED TYPE I
PORPHYRINOGENS OXIDIZE
INTO COLORED TYPE I
CEP H2O
A = ACETYL (2C) PORPHYRINS
P = PROPIONYL (3C)

X M = METHYL (1C)

THESE PORPHYRINS ARE

OVERPRODUCED BECAUSE
HEME FEEDBACK ON ALA
SYNTHASE IS REDUCED

4CO2
4CO2 REDUCED HEME
PRODUCTION IN
ERYTHROBLASTS
CAUSES ANEMIA
 CLINICAL CORRELATION
ACUTE INTERMITTENT PORPHYRIA (AIP)
ALSO KNOWN AS SWEDISH PORPHYRIA

MAIN CLINICAL MANIFESTATIONS:

ACUTE INTERMITTENT ATTACKS OF
NEUROVISCERAL SYMPTOMS WITH
WINE-COLORED URINE

ACUTE ATTACKS CAN BE PRECIPITATED

BY MANY FACTORS

INHERITANCE IS AUTOSOMAL DOMINANT

HETEROZYGOTES HAVE ABOUT 50% OF

L) NORMAL URINE
R) URINE FROM AN AIP
PATIENT DURING AN ATTACK
NORMAL ENZYME ACTIVITY
BUT MOST HETEROZYGOTES
NEVER DISPLAY SYMPTOMS (!)
 AIP IS CAUSED BY PARTIAL PORPHOBILINOGEN DEAMINASE DEFICIENCY

IN AIP THE URINE HAS VERY HIGH PBG

LEVELS DURING ACUTE ATTACKS
2H20

THE PBG REACTS SPONTANEOUSLY TO

FORM PORPHYRINS & OTHER COLORED
METABOLITES
AIP Nonenzymatic
4NH3

PORPHYRINS ETC.

A = ACETYL (2C)
P = PROPIONYL (3C)
 CLINICAL CORRELATION
VARIEGATE PORPHYRIA (VP)

MAIN CLINICAL MANIFESTATIONS:

ACUTE INTERMITTENT ATTACKS OF

NEUROVISCERAL SYMPTOMS
WITH PORT-WINE URINE (LIKE AIP)

PHOTODERMATITIS (UNLIKE AIP)

AUTOSOMAL DOMINANT INHERITANCE

MOST HETEROZYGOTES NEVER SHOW
SYMPTOMS (LIKE AIP)
HAND OF A VP
PATIENT
VP IS CAUSED BY PARTIAL PROTOPORPHYRINOGEN OXIDASE DEFICIENCY

IN VP THE URINE HAS HIGH PORPHYRIN III LEVELS

DURING ACUTE ATTACKS
COPROPORPHYRIN III > UROPORPHYRIN III

FECES ARE COLORED PURPLE MAINLY BY

PROTOPORPHYRIN IX THAT FORMS
SPONTANEOUSLY IN THE BLOOD FROM THE HIGH
LEVELS OF PROTOPORPHYRINOGEN IX

PROTOPORPHYRIN IX IS LESS WATER-SOLUBLE & IS

EXCRETED MAINLY IN THE BILE
BLOOD ALA & PB LEVELS ARE ALSO ELEVATED &
PRODUCE NEUROVISCERAL SYMPTOMS
3H2O2

O2 2H +
2CO2
 PORPHYRIA IS A DISEASE THAT MAY HAVE ALTERED HISTORY
KING GEORGE III OF GREAT BRITAIN
SUFFERED INTERMITTENT ATTACKS OF
BEHAVIORAL CHANGES (MADNESS)
ACCOMPANIED BY ABDOMINAL PAIN,
CONSTIPATION, SKIN RASHES & LIMB WEAKNESS

HIS PHYSICIANS REPORTED URINE &

STOOLS THE COLOR OF PORT WINE

MOST HISTORIANS NOW BELIEVE THAT

KING GEORGE III SUFFERED FROM
VARIEGATE PORPHYRIA
HETEROZYGOUS DEFICIENCY OF
PROTOPORPHYINOGEN OXIDASE

HISTORIANS ALSO BELIEVE THAT

HIS FITS OF MADNESS INFLUENCED
BRITAINS MISCONDUCT OF THE AMERICAN
GEORGE III REVOLUTION (1775 1783)
KING OF GREAT BRITAIN
1760 - 1820
 2. HEME DEGRADATION
HEME IS DEGRADED TO BILIRUBIN MAINLY IN THE SPLEEN

*
*

HEME OXYGENASE FORMS BILIVERDIN WITH

LOSS OF 1 CARBON AS CARBON MONOXIDE
THIS BREAKS THE PORPHYRIN RING
 BILIVERDIN IS THEN REDUCED TO BILIRUBIN BY
BILIVERDIN REDUCTASE

THIS IS AN NADPH-DEPENDENT REDUCTION THAT

ALSO OCCURS IN THE SPLEEN

UNCONJUGATED BILIRUBIN
IS EXPORTED TO THE PLASMA

UNCONJUGATED BILIRUBIN HAS LOW WATER-

SOLUBILITY & MUST BE TRANSPORTED IN PLASMA
BOUND TO ALBUMIN
ALBUMIN-BOUND UNCONJUGATED BILIRUBIN GOES TO THE LIVER
INSIDE THE HEPATOCYTE THE BILIRUBIN IS BOUND TO LIGANDIN

INSIDE THE HEPATOCYTE

UDP-GLUCURONYL TRANSFERASES
(UGT I & II) JOIN GLUCURONIC ACID
TO THE 2 PROPRIONATE GROUPS

THIS IS BILIRUBIN DIGLUCURONIDE OR

CONJUGATED BILIRUBIN

CONJUGATED BILIRUBIN IS MUCH

MORE WATER-SOLUBLE THAN
UNCONJUGATED BILIRUBIN
 CONJUGATED BILIRUBIN IS SECRETED INTO
THE BILE DUCTS VIA AN ABC-TRANSPORTER
FROM THERE IT MOVES INTO THE SMALL INTESTINE
INTESTINAL BACTERIA DECONJUGATE & REDUCE
THE BILIRUBIN TO FORM COLORLESS UROBILINOGENS

SOME OF THE
INTESTINAL
UROBILINOGEN IS
REABSORBED BACK
INTO THE BLOOD & IS
RE-EXCRETED IN THE
BILE & URINE

THE UROBILINOGENS
REMAINING IN THE
FECES ARE OXIDIZED
TO BROWN UROBILINS

UROBILINS CONTRIBUTE TO THE COLOR OF THE URINE

UROBILINS SUCH AS STERCOBILIN PRODUCE

MOST OF THE COLOR OF THE FECES
 CLINICAL CORRELATION
PHYSIOLOGICAL NEONATAL JAUNDICE
THIS IS CAUSED BY TRANSIENT DEFICIENCY OF
LIVER UDP-GLUCURONYLTRANSFERASES I & II
THE LIVER BEGINS TO
EXPRESS THE UGTs LATE
IN FETAL LIFE
IN MANY NEWBORN
X LIVERS UGTs ARE NOT
YET FULLY EXPRESSED

THERE IS MILD TO MODERATE

UNCONJUGATED
HYPERBILIRUBINEMIA

JAUNDICE SPONTANEOUSLY
DISAPPEARS IN A FEW DAYS AS THE
NORMAL LIVER MATURES

SERIOUS CLINICAL COMPLICATIONS NEWBORN INFANT

ARE VERY RARE WITH PNJ
 REVERSAL OF HYPERBILIRUBINEMIA CAN BE ACCELERATED BY PUTTING THE
NEWBORN UNDER BRIGHT LIGHTS (BILI LIGHTS) FOR A FEW HOURS EACH DAY

PHOTOTHERAPY
CONVERTS THE 2
BILIRUBIN DOUBLE
BONDS FROM
X TRANS TO CIS

THE CIS ISOMER IS MORE

POLAR & CAN BE
SECRETED BY THE LIVER
WITHOUT CONJUGATION

NEONATAL
PHOTOTHERAPY UNIT
 CLINICAL CORRELATION
CRIGLER-NAJJAR SYNDROME (CNS)
CNS IS CAUSED BY INHERITED HOMOZYGOUS DEFICIENCY
OF LIVER UDP-GLUCURONYLTRANSFERASE I
LIVER UGT II ALONE
CANNOT HANDLE ALL OF
THE BILIRUBIN

X NEWBORNS HAVE SEVERE

UNCONJUGATED
HYPERBILIRUBINEMIA

SEVERE HYPERBILIRUBINEMIA CAN LEAD TO

KERNICTERUS (DEPOSIT OF BILIRUBIN IN THE
BRAIN) WITH DEVELOPMENT OF ENCEPHALOPATHY

CNS PATIENTS USUALLY DIE IN INFANCY

EARLY LIVER TRANSPLANTATION

HAS SAVED SOME PATIENTS
END OF SIM 3-1