Biosynthesis of Porphyrin

K.H.Timotius
June 2014

Outline
1. Structure of porphyrin
2. Biosynthesis of heme
3. Porphyria

1. Chemistry of porphyrin
porphyrin,any of a class of water-soluble, nitrogenous
biological pigments (biochromes), derivatives of which
include the hemoproteins (porphyrins combined with
metals and protein).
Examples of hemoproteins are
a. the green, photosynthetic chlorophylls of higher plants;
b. the hemoglobins in the blood of many animals;
c. the cytochromes, enzymes that occur in minute
quantities in most cells and are involved in oxidative
processes; and
d. catalase, also a widely distributed enzyme that
accelerates the breakdown of hydrogen peroxide.

1. Chemistry of porphyrin
Porphyrins have complex cyclic structures.
All porphyrin compounds absorb light intensely
at or close to 410 nanometres.
Structurally, porphyrin consists of four pyrrole
rings (five-membered closed structures
containing one nitrogen and four carbon atoms)
linked to each other by methine groups (CH=).
The iron atom is kept in the centre of the
porphyrin ring by interaction with the four
nitrogen atoms.

Spectrometry UV-VIS

2. Synthesis of Porphobilinogen and

heme
The first reaction in heme biosynthesis takes
place in the mitochondrion and involves the
condensation of one glycine and one
succinylCoA by the pyridoxal phosphatecontaining enzyme, -aminolevulinic acid
synthase (ALAS).
Delta-aminolevulinic acid (ALA) is also called 5aminolevulinic acid.
This reaction is both the rate-limiting reaction
of heme biosynthesis, and the most highly
regulated reaction

hemoprotein
Hemoglobin: transport oksigen dalam
darah
Mioglobin: penyimpanan oksigen di otot
Sitokrom c: terlibat dalam rantai transpor
elektron
Sitokrom P450: hidroksilasi xenobiotik
Katalase: penguraian hidrogen peroksida
Triptofan pirolase: oksidasi triptofan
Vitamin B12: ?

2. Synthesis of Porphobilinogen and

heme
Two forms of ALAS: ALAS1 and ALAS2.
ALAS1 is considered a house-keeping gene and is expressed in
all cells.
ALAS2 is an erythroid-specific form of the enzyme and is
expressed only in fetal liver and adult bone marrow.
The ALAS1 gene is located on chromosome 3, whereas the
ALAS2 gene is located on the X chromosome.
Deficiencies in ALAS2 result a disorder called X-linked
sideroblastic anemia, XLSA. Sideroblasts are erythroblasts with
non-heme iron-containing organelles, called siderosomes. XLSA
has also been called congenital sideroblastic anemia,
hereditary sideroblastic anemia, hereditary iron-loading
anemia, X-linked hypochromic anemia, hereditary hypochromic
anemia, and hereditary anemia.

2. Synthesis of Porphobilinogen and

heme
Following synthesis mitochondrial ALA is
transported to the cytosol, where ALA
dehydratase (also called porphobilinogen
synthase) dimerizes two molecules of ALA to
produce the pyrrole ring compound
porphobilinogen.
The next step in the pathway involves the
head-to-tail condensation of four molecules of
porphobilinogen to produce the linear
tetrapyrrole intermediate,
hydroxymethylbilane.

2. Synthesis of Porphobilinogen and

heme
The enzyme for this condensation is porphobilinogen
deaminase (PBG deaminase).
This enzyme is also called hydroxymethylbilane
synthase or uroporphyrinogen I synthase.
Hydroxymethylbilane has two main fates.
The most important is regulated, enzymatic conversion
to uroporphyrinogen III, the next intermediate on the
path to heme.
This step is mediated by a holoenzyme comprised of
uroporphyrinogen synthase plus a protein known as
uroporphyrinogen III cosynthase. Hydroxymethylbilane
can also non-enzymatically cyclize forming
uroporphyrinogen I.

3. Regulation of Heme
Biosynthesis
Although heme is synthesized in
virtually all tissues, the principal sites
of synthesis are erythroid cells (85%)
and hepatocytes (accounting for
nearly all the rest of heme synthesis).
The differences in these two tissues
and their needs for heme result in
quite different mechanisms for
regulation of heme biosynthesis.

3. Regulation of Heme
Biosynthesis
In hepatocytes, heme is required for incorporation into the
cytochromes, in particular, the P450 class of cytochromes
that are important for detoxification. In addition numerous
cytochromes of the oxidative-phosphorylation pathway
contain heme.
The rate-limiting step in hepatic heme biosynthesis occurs
at the ALA synthase catalyzed step, which is the committed
step in heme synthesis. Heme itself functions as a corepressor in the inhibition of ALA synthase gene expression.
The Fe3+ oxidation product of heme is termed hemin.
Hemin acts as a feed-back inhibitor on ALA synthase. Hemin
also inhibits transport of ALA synthase from the cytosol (its'
site of synthesis) into the mitochondria (its' site of action).

3. Regulation of Heme
Biosynthesis
In erythroid cells all of the heme is synthesized for incorporation
into hemoglobin and occurs only upon differentiation when
synthesis of hemoglobin proceeds.
When red cells mature both heme and hemoglobin synthesis
ceases. The heme (and hemoglobin) must, therefore, survive for
the life of the erythrocyte (normally this is 120 days). In
reticulocytes (immature erythrocytes) heme stimulates protein
synthesis. The mechanism of this mode of heme-mediate
regulation of protein synthesis is described in the
Protein Synthesis section. Additionally, control of heme
biosynthesis in erythrocytes occurs at numerous sites other than
at the level of ALA synthase. Control has been shown to be
exerted on ferrochelatase, the enzyme responsible for iron
insertion into protoporphyrin IX, and on porphobilinogen
deaminase.

5. Clinical Aspect of Heme

Metabolism
two types: Porphyrias and Bilirubinemias.
1. Disorders that arise from defects in the
enzymes of heme biosynthesis are
termed the porphyrias and cause
elevations in the serum and urine content
of intermediates in heme synthesis.
2. Inherited disorders in bilirubin
metabolism lead to hyperbilirubinemia.

Porphyrias
are both inherited and acquired disorders in heme synthesis.
These disorders are classified as either erythroid or hepatic,
depending upon the principal site of expression of the enzyme defect.
Eight different porphyrias have been classified encompassing defects
in each of the enzymes of heme synthesis.
Defects in hepatic uroporhyrinogen decarboxylase (UROD) results in
type I porphyria cutanea tarda (PCT I), whereas deficiencies in the
non-hepatic forms of UROD result in type II PCT (PCT II). PCT is the
most commonly occurring type of porphyria.
It should be noted that no porphyria has been identified resulting from
defects in the house-keeping form of ALAS (ALAS1).
The most commonly occurring hepatic porphyria is acute intermittent
porphyria, AIP which is caused by a defect in porphobilinogen
deaminase, (PBG deaminase). This enzyme is also called
hydroxymethylbilane synthase or rarely uroporphyrinogen I synthase.

Porphyrias
All of the porphyrias lead to excretion of heme
biosynthetic byproducts that turn the urine red and
when deposited in the teeth turn them reddish
brown.
Accumulation of these byproducts in the skin renders
it extremely sensitive to sunlight causing ulceration
and disfiguring scars.
Increased hair growth (hypertrichosis) is also a
symptom of the porphryias leading to appearance of
fine hairs over the entire face and on the extremities.
This latter symptom lends to the description of
"werewolf syndrome" in many porphyria patients.

Pustaka
Murray, R.K., D.K.Granner, and
V.W.Rodwell (Eds.), 2009. Biokimia
Harper. Edisi 27. Bab 31. Porfirin dan
Pigmen Empedu (Robert K. Murray)
Halaman 288- 303. Penerbit Buku
Kedokteran, Jakarta