HEME SYNTHESIS

G HARIPRASATH
RESEARCH SCHOLAR
DEPT OF BIOCHEMISTRY
RMMC
ANNAMALAI UNIVERSITY
 HEME SYNTHESIS
Heme Metallo HC CH
porphyrin.
Porphyrins Cyclic
compounds formed by
4 pyrrole rings linked
HC CH
through methyne or N
methenyl bridges H
(=CH-)
Porphyrins usually
forms complexes with
metal ions.
Pyrrole ring
 Hemeis the prosthetic group of hemoglobin, myoglobin,
and thecytochromes.
It is asymmetric.
Porphyrin of heme Protoporphyrin.
One ferrous (Fe2+) atom in the center of the tetrapyrrole
ring.
So heme is ferro protoporphyrin.
 Acetate, Propionyl, Methyl and Vinyl groups are often
attached as side chains and depending on their positions
they form different isomers.
Important porphyrins: Uroporphyrins, Protoporphyrins
and Coproporphyrins.
Protoporphyrins has Methyl, Propionate and Vinyl side
chains.
Uroporphyrins has acetate and propionate chains.

Coproporphyrins has Methyl and propionate chains.

Type III porphyrins are normally important for humans.

 Heme is also the Prosthetic group for
Myoglobin
The Cytochromes
Catalase and Tryptophan pyrrolase
Nitric Oxide Synthase
Magnesium porphyrin chlorophyll

Porphyrins - used in cancer phototherapy. Upon

administration taken up more by cancer tissues.
Then the patient is exposed to argon laser
which excites porphyrins producing cytotoxic
effects. Photodynamic therapy.
BIOSYNTHESIS OF HEME
Site All tissues in the body except mature
erythrocytes (lacks Mitochondria).
85% in erythroid precursor cells in bone marrow.
Under the control of erythropoietin.
Remaining mainly in hepatocytes.

Organelle Partly cytoplasmic & Partly

mitochondrial.
 STEPS
Synthesis of Porphobilinogen (Monopyrrole)
Synthesis of Uroporphyrinogen (Tetrapyrrole)

Conversion of Uroporphyrinogen to
Protoporphyrin.
Formation of heme
 SYNTHESIS OF
PORPHOBILINOGEN
 Hemesynthesisbegins with condensation of glycine & succinyl-
CoA, with decarboxylation, to formd-aminolevulinic acid(ALA).
1. ALA synthesis Mitochondria.

Rate limiting step.

ALA synthase requires pyridoxal phosphate & Mg++ for its

activity.
d-Aminolevulinate Synthase(ALA Synthase) is therate

limiting stepof the heme synthesis pathway.

Heme feed back inhibitor.

Hemin produced by oxidation of heme. Not taken up by protein.

Inhibits ALA synthesis.

2. Phorphobilinogen synthesis Cytosol.

PBGSynthase(Porphobilinogen Synthase), also called ALA

Dehydratase, catalyzes condensation oftwomolecules ofd-

aminolevulinic acid (ALA) to formporphobilinogen(PBG).
ALA dehydratase contains zinc.

Sensitive to inhibition by lead lead poisoning increased ALA &

anemia.
FORMATION OF UROPORPHYRINOGEN III BY
CONDENSATION OF FOUR
PORPHOBILINOGENS
 Four PBG molecules condense
Catalysed by PBG deaminase or HMB synthase.

Four molecule of PBG condense in a head to tail manner to

form Hydroxy methylbilane (HMB).
Uroporphyrinogen III synthase: Converts HMB to
Uroporphyrinogen III.
Normal condition Uroporphyrinogen III gets formed.

Under certain porphyrias HMB cyclises spontaneously to

form Uroporphyrinogen I.
CONVERSION OF UROPORPHYRINOGEN
TO PROTOPORPHYRIN
 Enzyme Uroporphyrinogen decarboxylase.
Four Acetate groups of Uroporphyrinogen III were

decarboxylated to Methyl groups to form

coproporphyrinogen III.
Site cytosol.

Conversion of Coproporphyrinogen III to

Protoporphyrinogen IX:
Enzyme Coproporphyrinogen oxidase.

Oxidative decarboxylation.

Protoporphyrinogen IX is converted to protoporphyrin IX

by protoporphyrinogen oxidase
Oxidation occurs by molecular oxygen.

6H+ are liberated.

HEME FORMATION
 By incorporation of iron (Fe 2+).
Partly spontaneous.
Enhanced by Ferrochelatase (heme synthetase).
May be inhibited by lead.
Insertion of an atom of Fe++ into central position
of protoporphyrin IX.
Heme produced can be then coupled to various
proteins-conjugated proteins: hemoglobin,
myoglobin, cytochrome C, catalases and
peroxidases.
 REGULATION OF HEME SYNTHESIS

Regulator enzyme ALA synthase & Hydroxy methyl bilane

synthase.
Heme negative regulator.
Absence of heme: Synthesis of ALA synthase & HMB synthase
increases greatly.
Enzyme synthesis, as well as its transport to the mitochondria, is
also inhibited by elevated levels hemin (oxidised product of
heme).
Many xenobiotics on administration increases ALA synthase
activity .Eg., barbiturates-phenobarbitol and griseofulvin These
drugs are usually metabolized by the microsomal cytochrome
P450 mono-oxygenase system. Which diminishes intracellular
heme conc.
ALA synthase Requires pyridoxal phosphate (Vitamin B6) as a
coenzyme.
Bio availability of iron too plays an important role.
 PORPHYRIAS
Defect in heme biosynthesis accumulation of porphyrins or
its precursors porphyrias.
Can be hereditary or acquired (common).

Classification:

Hepatic

Erythropoietic.

Hepatic forms neurological symptoms

Erythropoietic forms cutaneous symptoms.

Salient features:

Occurrence: Mutation of any gene encoding enzymes of heme

synthesis.
Charectiresed by non synthesis of metabolic product beyond

the point of enzyme defect. Accumulation of intermediates

prior to block.
All Porphyrias are autosomal dominant. Exception

Congenital erythropoietic porphyria (autosomal

recessive).
1. Enzyme defect earlier in the pathway ALA, PBG
and HMB accumulates. Causes Abdominal pain
and neuropsychiatric symptoms.
2. Enzyme defect later in the pathway
porphyrinogens are accumulated. Upon oxidation
they produces corresponding porphyrins which
cause photo sensitivity resulting in skin injury.
(generation of free radicals which damage cellular
component).
3. Consumption of antioxidants may benefit.
4. Treatment: gene replacement therapy
(experimental).
5. Administration of heme or hematin.
6. Avoiding drugs that induce cyt P450.
TYPES OF PORPHYRIAS
ALA synthase deficiency: mutation in gene
encoding ALA synthase.
No accumulation of porphyrins.

Sideroblastic anemia.

May respond to supplementation of pyridoxine.

Plumboporphyria: Deficiency of PBG synthase

or ALA dehydratase. Occurrence (rare) Lead
poisoning.
PORPHYRIAS ARE RESPONSIBLE FOR THE
ENZYME DEFECTS
 Acute interittent porphyria:
Enzyme defect: HMB synthase.

Products accumulated: Porphobilinogen & ALA.

Urine colourless but darkens on exposure to air due to

photooxidation.
Abdominal pain and neuropsychiatric symptoms.

Psychiatric symptoms reduced activity of tryptophan

pyrrolase.
Administration of intravenous hematin may ameliorate
the symptoms.
Administration of Barbiturates, ethanol, steroids can
precipiate the manifestations.
No photosensitivity.
 Congenital erythropoietic porphyria:
Gunthers disease.

Autosomal recessive.

Enzyme defect: uroporphyrinogen III synthase

(low activity).
Diversion of pathway. Type I isomers gets formed
(Uroporphyin I and Coproporphyrin I).
Urine dark red in colour.

High photosensitivity is observed.

Teeth exhibit fluoresence when exposed to UV

light (erythrodontia).
 Porphyria Cutanea tarda: Most common.
Enzyme defect: partial deficiency of uroporphyrinogen
decarboxylase.
Precipitated by excess alcohol consumption and iron
overload.
Uroporphyrins I and III are excreted in urine.

Photosensitivity is observed.

Hereditary coproporphyria:

Enzyme defect: Coproporphyrinogen oxidase.

Coproporphyrinogen III and other metabolites of heme

synthesis prior to enzyme block accumulates.
Photosensitivity and neurological attacks.
 Variegate Porphyria:
Enzyme defect protoporphyrin oxidase.

Products accumulate protoporphyrin, coproporphyrin,

uroporphyrin, PBG.
Neurological attacks and photosensitivity are observed.

Erythropoietic Protoporphyria:

Enzyme defect ferrochelatase.

Elevated conc of protoporphyrin in blood, bile and feces.

Protoporphyrin strongly hydrophobic.

Mild photosensitivity.

No neurologic symptoms.

May exhibit cholelithiasis (gall stone disease) and portal

hypertension.
ACQUIRED PORPHYRIAS
Secondary to CKD, few malignancies, liver
disease.
Exposure to heavy metals Pb, Hg, As.

Toxic porphyrias.

Associated with anemia.