HEME SYNTHESIS AND PORPHYRIAS

HEME PROTEIN
o Heme is ferroprotoporphyrin IX, it is the prosthetic group of a number of
heme proteins
o Heme proteins:
Mb, Hb, mitochondrial chromosomes, microsomal cytochrome,
catalase and peroxidase, tryptophan pyrrolase
o Heme is usually associate noncovalently with its apoprotein through the
coordination of its iron with an aa side chain N
The heme in cytochrome C derivatives are an exception in being
covalently bound to the apoprotein through thioether linkages
STRUCTURE OF PORPHYRINS
o Heme is the iron chelate of protoporphyrin IX
o Porphyrins-metal free cyclic tetrapyrroles which are named according to
the nature and arrangement of their side chain groups
Positions 5,10,15,20 are C atoms that serve as bridges bt the 4
pyrrole rings (aka alpha, beta, gamma, delta)
These C bridges are unsaturated in porphyrins and are
called methene groups (-CH=)
o Because they are unsaturated, the entire structure is
conjugated, therefore, porphyrins are colored (usu a
shade or red or brown)
o Porphyrin Biosynthesis
First the molecule goes through a stage where the bridge carbons
are saturated=methylene bridge
Its presence in the molecule disrupts the conjugation of the
molecule and therefore, the tetrapyrrole is colorless
Porphyrinogen=when all the carbon bridges are methylene
groups
Poryphrins are named by the groups attached to them
Acetic Acid
Proprionic acid
Methyl
Vinyl
Occur as:
o AP, MP, MV
The porphyrins of interest to us contain 4 pyrroles in the
combinations below:
o Uropophyrin- 4AP (most soluble)
o Coproporphyrin-4 MP
o Protoporphyrin-2MP, 2MV (least soluble)

It is possible to draw a number of isomers of these

porphyrins depending on how different the pyrroles are put
together
o Type III is the naturally occurring isomer or
uroporyphyrin and coproporphyrin (APAPAPPA
and MPMPMPPM respectively)
o Protoporphyrin IX is the naturally occurring isomer
(MVMVMPPM)
These compounds can occur with methane or methylene
bridges
PROPERTIES OF PORPHYRINS
o Color
usually a shade of red or brown with a distinctive light absorption
spectrum
major absorption is at 400nm (the Soret band) with several
smaller bands between 500-630nm
metal-free porphyrins emit an intense red fluorescence
when excited by light at ~400nm, but heme does not
Porphyrin-sensitized injury aka photosensitivity-the
fluorescence in vivo of porphyrins in the presence of O damages
cells
Based on injury to plasma and lysosomal membranes
Photosensitization occurs when abnormally large quantities
of porphyrins are present either by experimentation or by a
derangement in porphyrins synthesis called porphyria
o Symptoms=itching, edema, erythema and
ulceration
Another problem due to elevated porphyrins levels is in regards to
water solubility
In general, porphyrins solubility increases as the number of
carboxylic acid side chains increases
o Uro and coproporphyrins can be excreted in urine
whereas protoporphyrin is so insoluble it must be
secreted in feces via bile
THE PATHWAY OF HEME SYNTHESIS
o The cells of most tissues can synthesize heme compounds but the liver and
bone marrow produce the most
In bone marrow, heme is inc. into Hb, the liver normally produces
15% of the total heme synthesized in the body and the bone
marrow does the rest
In rats, 65% of heme synthesized in the liver is used for the
formation of microsomal cytochrome P450 and 6% for
mitochondrial cytochromes

o The porphyrins ring is synthesized entirely from glycine and succinate (as
succinyl CoA)
2 key intermediates=aminolevulini acid ALA and porphobilinogen
PBG
o #1-8 enzymes are involved from glycine; the first and last three are in the
mitochondria requiring the cooperation of the mitochondria and cytostolic
compartments for heme synthesis
ALA IS FORMED IN THE FIRST STEP
PBG (the pyrrole precursor of heme) IS FORMED IN
THE SECOND
o 4 molecules of PBG are condensed in the
pathway to heme
o #2-ALA synthase, the first enzyme of the pathway, is dependent on
pyridoxal phosphate for activity
This coenzyme functions to form a Schiff base with the glycine
before is reats with succinyl CoA
o #3-ALA is the initial intermediate=committed to heme synthesis and
rate controlling step!
o #4-from PBG until protoporphyrin IX, the precursors of heme are
synthesized reactions involving reduced ridge Cs of the porphyrinogen
type
Only the final enzyme, ferrochelatase, uses a porphyrins instead of
a porphyrinogen
o #5-Reduced iron (Fe2+) is incorporated into heme by ferrochelatase
o #6-PBG deaminase catalyzes the condensation of PBG molecules to form
a linear tetrapyrrole
When PBG acts in the presence of a 2nd enzyme, uroporphyrinogen
III cosynthase, proper closure of the tetrapyrrole occurs
In the absence of this enzyme, type I is formed which is unusable
o #7-uroporphyrinogen III is the first porphyrinogen formed in the
biosynthesis of heme
It is converted to protoporyphyrin IX by enzymatic reaction of its
side chains
4 acetic groups methly groups
2 proprionic acid groupsvinyl groups
REGULATION OF HEME BIOSYNTHESIS
o In liver:
ALA synthase (the 1st enzyme of the pathway) catalyzes the ratelimiting step and is the major point of regulation of the pathway in
the liver (but not in erythroid cells)
level of ALA synthase is regulated by the concentration of
free heme, the product of the pathway, and the
autooxidation product hemin (heme with Fe+3)
o There are 3 mechanisms which might regulate the rate of heme synthesis

1. Feedback inhibition of the isolated enzyme by hemin

Does not seem to be physiologically important
2. Hemin represses the synthesis of ALA synthase
3. Hemin inhibits the transport of cytosolic ALA synthase into the
mitochondria
THEREFORE, HIGH LEVELS OF HEME PREVENT THE
SYNTHESIS OF HEME!
o Many compounds increase ALA synthase and result in overproduction of
liver ALA, PBF and porphyrins
Ie. Phenobarbital-related barbiturates, naturally occurring
steroids (including estrogens) and other drugs
o There are tissue-specific ALA synthase isozymes which are encoded by 2
separate genes
ALAS1-housekeeping ALA synthase gene expressed in liver and
non-erythyroid tissues
ALAS2-expressed only in erythroid cells
o In erythroid cells:
The major regulation of heme synthesis in these cells is no via the
repression of ALA synthesis by heme as in the liver
In fact, hemin stimulates heme formation in several types of
erythroid cells
There is an erythroid-specific ALA synthase gene which has a
distinct mode of regulation tied to cell differentiation
ALAS-E is not inducible by the drugs that induce ALAS1
PORPHYRIA
o A group of related but distinct diseases, they are inherited and acquired
diseases characterized by:
Defects in specific enzymes in heme biosynthesis
Increased accumulation and excretion of intermediates of the
pathway
Most dramatic clinical symptoms=neurological disorders and
cutaneous photosensitivity
Excess ALA and PBF=peripheral neuritis, abdominal pain
and intestinal spasms
Excess porphyrins in plasma(rather than in blood
cells)=skin lesions
o Porphyrins transported by albumin
CLASSIFICATION OF PORPHYRIAS
o Classified as erythropoietic or hepatic, symptoms are essentially the same
bc overproduced porphyrins can enter plasma from both sources
COMMENTS ON PORPHYRIAS
o Erythropoeitic porphyrias
Congenital erythropoietic porphyria (CEP) is very rare
Severe photosensitivity, red cell hemolysis

Erythropoietic porphyria protoporphyria (EPP)

Usually exhibits milder clinical symptoms with little or no
hemolysis
o Hepatic Porphyrias
5-aminolevulinic acid dehydratase-deficient porphyria (ADP),
acute intermittent porphyria (AIP), porphyria cutanea tarta (PCT),
hereditary coproporphyria (HCP) and variegate porphyria (VP)
Variable degree of clinical expression
Hormonal, drug and nutritional factors predispose patients
to full expression of the disease
o These agents precipitate clinical expression in
asymptomatic patients carrying the genetic defect
o Most act by inducing higher levels of ALA synthase
o PCT is fairly common
More frequent in males over 40 who drink
and women on birth control
o ADP is extremely rare!
One would expect in some cases the accumulation of the gens in
the urine, but porphyrinogens oxidize spontaneously to the
porphyrins form during and after excretion; thus, a urine specimen
will gradually darken if left standing
The enzymatic defect in various porphyrias is genetic
Enzymatic activity is REDUCED
LEAD POISONING
o Usually occurs from the ingestion of peeling leaded paints or other
exposure
o Symptoms are similar to those AIP
o Lead tends to concentrate in erythroid cells
o Erythrocyte ALA dehydratase and ferrochelatase are particularly sensitive,
resulting in a kind of acquired porphyria
o Inhibits erythrocyte heme synthesis more than liver heme synthesis
o Excess excretion of ALA and proto IX and Copro III
o Possible photosensitiviy and noticeable anemia
o Abodominal and neurological problems