Porphyrins & Bile Pigments

FY WIDODO
University of Wijaya Kusuma Surabaya
Medical Faculty
Department of Biochemistry

TUGAS KULIAH
PORFIRIN/PURIN/PIRIMIDIN
MEMBUAT RANGKUMAN KULIAH
MANFAAT YANG BISA DIAMBIL DARI MATERI KULIAH
INI (BERSIFAT KLINIS)
MINIMUM 3 HALAMAN FOLIO, MAKSIMUM 5
HALAMAN FOLIO UNTUK 2 MATA KULIAH, PORFIRIN
DAN PURIN/PIRIMIDIN
TULISAN TANGAN !!!
TULIS NAMA, NPM, MATERI KULIAH, NAMA DOSEN
DISERAHKAN PALING LAMBAT 1 MINGGU SETELAH
SELESAI KULIAH PORFIRIN/PURIN / PIRIMIDIN

PORFIRIN
Cyclic compounds formed by the linkage
of four pyrrole rings through methyne (
HC =) bridges
Formation of complexes with metal ions
bound to the nitrogen atom of the pyrrole
rings
Examples of Some Important Human Hemoproteins
Protein

Function

Hemoglobin

Transport of oxygen in blood

Myoglobin

Storage of oxygen in muscle

Cytochrome c

Involvement in electron transport

chain

Cytochrome P450

Hydroxylation of xenobiotics

Catalase

Degradation of hydrogen peroxide

Tryptophan

pyrrolase

Oxidation of tryptophan

A (acetate)
P (propionate)
M (methyl)

Addition of iron to protoporphyrin

to form heme. V (vinyl) = CHCH2.

Formation of Heme

The two starting materials are succinyl-CoA and glycine

Pyridoxal phosphate "activate" glycine
Condensation reaction between succinyl-CoA and glycine -amino-ketoadipic acid
-aminolevulinate (ALA)
decarboxilation

Enzyme: ALA synthase

MITOCHONDRIA

Two molecules of ALA are condensed by the enzyme ALA

dehydratase to form two molecules of water and one of
porphobilinogen (PBG)
ALA dehydratase is a zinc-containing enzyme and is
sensitive to inhibition by lead, as can occur in lead
poisoning.
CYTOSOL

The formation of a cyclic tetrapyrroleie, a porphyrinoccurs by

condensation of four molecules of PBG

These four molecules condense in a head-to-tail manner to form a

linear tetrapyrrole, hydroxymethylbilane (HMB)

Enzyme: uroporphyrinogen I synthase = porphobilinogen (PBG)

deaminase = HMB synthase
cyclizes spontaneously uroporphyrinogen I

HMB
uroporphyrinogen III synthase uroporphyrinogen III

These compounds are colorless (porphyrinogens)

auto-oxidized porphyrins (colored).

 uroporphyrinogen I uroporphyrinogen decarboxylase

coproporphyrinogen I
uroporphyrinogen III coproporphyrinogen III
Acetate (A) methyle (M) : decarboxylated by
uroporphyrinogen decarboxylase
cytosol
coproporphyrinogen III coproporphyrinogen oxidase
protoporphyrinogen III
protoporphyrinogen III protoporphyrinogen oxidase
protoporphyrin III
Requires molecular oxygen.
Protoporphyrin III + Fe2+ Heme
Enzyme: ferrochelatase (heme synthase)
mitochondria

Decarboxylation of uroporphyrinogens to coproporphyrinogens in

cytosol. (A, acetyl; M, methyl; P, propionyl.)

Heme biosynthesis occurs in most mammalian cells with the exception of

mature erythrocytes, which do not contain mitochondria.
However, approximately 85% of heme synthesis occurs in erythroid
precursor cellsin the bone marrow and the majority of the remainder in
hepatocytes.

Regulation in Biosynthesis of Heme

ALA synthase occurs in both hepatic (ALAS1) and erythroid (ALAS2)
forms
The rate-limiting enzyme: ALAS1 feedback inhibition
Heme (-) ALAS1
Heme (+) ALAS1
Aporepressor molecule + Heme negative regulator of the synthesis
of ALAS1
Drugs (eg, barbiturates, griseofulvin) increase in ALAS1.
Most of these drugs are metabolized by a system in the liver that
utilizes cytochrome P450
Glucose can prevent derepression of ALAS1 in liver as can the
administration of hematin (an oxidized form of heme).

PORPHYRIAS

Disorders due to abnormalities in the pathway of biosynthesis of

heme genetic or acquired

Enzyme Involved

Type, Class,

Major Signs and

Symptoms

Results of Laboratory
Tests

ALA synthase (erythroid

form)

X-linked sideroblastic
anemia3 (erythro-poietic)

Anemia

Red cell counts and

hemoglobin decreased

ALA dehydratase

ALA dehydratase deficiency (hepatic)

Abdominal pain,
neuropsychiatric
symptoms

Urinary ALA and

coproporphyrin III
increased

Uroporphyrinogen I
synthase

Acute intermittent
porphyria (hepatic)

Abdominal pain,
neuropsychiatric
symptoms

Urinary ALA and PBG

increased

Uroporphyrinogen III
synthase

Congenital erythropoietic
(erythropoietic)

No photosensitivity

Urinary, fecal, and red cell

uroporphyrin I increased

Uroporphyrinogen
decarboxylase

Porphyria cutanea tarda

(hepatic)

Photosensitivity

Urinary uroporphyrin I
increased

Coproporphyrinogen
oxidase

Hereditary
coproporphyria (hepatic)

Photosensitivity, abdominal pain, neuropsychiatric symptoms

Urinary ALA, PBG, and

coproporphyrin III and fecal
coproporphyrin III increase

Protoporphyrinogen
oxidase

Variegate porphyria
(hepatic)

Photosensitivity, abdominal pain, neuropsychiatric symptoms

Urinary ALA, PBG, and

coproporphyrin III and fecal
protoporphyrin IX
increased

Protoporphyrinogen
oxidase

Variegate porphyria
(hepatic) (MIM 176200)

Photosensitivity, abdominal pain, neuropsychiatric symptoms

Urinary ALA, PBG, and

coproporphyrin III and fecal
protoporphyrin IX
increased

 In general, the porphyrias described are inherited in an autosomal

dominant manner, with the exception of congenital erythropoietic
porphyria, which is inherited in a recessive mode
The signs and symptoms of porphyria result from either a deficiency of
metabolic products beyond the enzymatic block or from an
accumulation of metabolites behind the block.
If the enzyme lesion occurs early in the pathway, clinically, patients
complain of abdominal pain and neuropsychiatric symptoms relate
to elevated levels of ALA or PBG or to a deficiency of heme
If the enzyme blocks later in the pathway result in the accumulation of
the porphyrinogens, their oxidation products, the corresponding
porphyrin derivatives, cause photosensitivity, a reaction to visible light
of about 400 nm The porphyrins, when exposed to light of this
wavelength, are thought to become "excited" and then react with
molecular oxygen to form oxygen radicals injure lysosomes and
other organelles release their degradative enzymes skin
damage, including scarring.
The porphyrias can be classified on the basis of the organs or cells
that are most affected: erythropoietic, hepatic & erythrohepatic

Barbiturates, griseofulvin cytochrome P450

heme
ALAS1
porphyria
Diagnosis: clinical and family history, the physical examination, and
appropriate laboratory tests
Treatment: - symptomatic
- to avoid drugs that cause induction of cytochrome P450.
- glucose,hematin may repress ALAS1
- photosensitivity : carotene free radicals
Sunscreens

Katabolisme Heme
12 x 108 erythrocytes are destroyed per hour in 1 day turns over
approximately 6 g of hemoglobin
globin amino acids reused;
iron pool
Enzyme: complex enzyme system called heme oxygenase

METABOLISME BILIRUBIN
PENGAMBILAN BILIRUBIN OLEH HATI
Bilirubin hanya sedikit larut dalam plasma & terikat dengan albumin
Obat / antibiotika kompetisi untuk berikatan dg albumin
LIVER: Bilirubin dilepas dari albumin diambil pada permukaan
sinusoid hapatosit Sistem Transport Berfasilitas (facilitated transport
system = carrier-mediated saturable system) masuk ke sel hati
berikatan dengan cytosolic protein (ligandin, protein Y). Ikatan ini juga
menjaga agar bilirubin tidak kembali ke aliran darah lagi.
Aktivitas sistem ini menurun pada keadaan patologis

kuning

hijau

KONJUGASI BILIRUBIN
LIVER: Bilirubin yang non-polar polar
Konjugasi dengan GLUKORONAT Bilirubin diglukoronida
(conjugated, "direct-reacting" bilirubin) polar
Enzyme: glucuronosyltransferase (dlm retikulum endoplasma)

Donor glukuronosil: UDP-glucuronic acid

Enzim dapat diinduksi oleh Phenobarbital
Bilirubin diglukoronida diekskresi melalui faeces
when exist abnormally in human plasma (eg, in obstructive jaundice),
they are predominantly monoglucuronides
Obstructive jaundice bilirubin conjugates (predominan
monoglukuronida)

Bilirubin diglukoronida

METABOLISME BILIRUBIN DALAM USUS

Diglukoronida

Bilirubindiglukoronida

Bilirubin
glukoronidase

Bakteri usus

Bilirubin direduksi oleh flora usus UROBILINOGEN (tak berwarna)

Sebagian kecil urobilinogen diabsorbsi & diresekresi melalui liver (SIKLUS
ENTEROHEPATIK)
Sebagian besar urobilinogen oksidasi UROBILIN (kuning) faeces
Urobilinogen dlm urine abnormal

Diagrammatic representation of the three

major processes (uptake, conjugation,
and secretion) involved in the transfer of
bilirubin from blood to bile. Certain proteins
of hepatocytes, such as ligandin (a member
of the glutathione S-transferase family of
enzymes) and Y protein, bind intracellular
bilirubin and may prevent its efflux into the
blood stream. The process affected in a
number of conditions causing jaundice
is also shown.

HIPERBILIRUBINEMIA

Bilirubin dlm darah >1 mg/dl.

Penyebab: - produksi bilirubin tidak sebanding dg ekskresi hati
- kegagalan ekskresi hati karena:
- kerusakan hati
- obstruksi saluran ekskresi
Bilirubin masuk ke jaringan KUNING / ICTERUS / JAUNDICE
Ehrlichs test dari van den Bergh:
- reagen diazo + Bilirubin + etanol senyawa Azo (ungu-merah)
INDIRECT REACTION BILI UNCONJUGATED (FREE)
- reagen diazo + Bilirubin (tanpa etanol) senyawa Azo
DIRECT REACTION BILI CONJUGATED
Retention hyperbilirubinemia: unconjugated
Regurgitation hyperbilirubinemia : conjugated
Bili unconjugated dpt menembus blood-brain barrier encephalopathy
Kern icterus

Jaundice

UNCONJUGATED HYPERBILIRUBINEMIA
A.

B.

Hemolytic Anemias
Biasanya ringan (< 4 mg/dL; < 68.4 mol/L) kapasitas hati besar
utk mengelola bilirubin (uptake, konjugasi, ekskresi)

Neonatal Physiologic Jaundice

- Hemolisis > metabolisme bilirubin dlm hati
- aktivitas / sintesis UDP-glukoronosil transferase
- Dpt menyebabkan Kern icterus
- Tx: Phenobarbital +

C. Crigler-Najjar Syndrome, Type I

- = Congenital nonhemolytic jaundice
- Autosomal resesif; mutasi pada gen pengkode enzim BilirubinUGT UDP-glukoronosil transferase
- Bilirubin bisa sd > 20 mg/dL bilirubin tdk bisa dikonjugasikan
- Klinis: ikterus kongenital berat fatal dalam 15 bulan
- Tx:
, liver transplant. Phenobarbital tdk menolong.
D. Crigler-Najjar Syndrome, Type II
- Mutasi pada gen pengkode enzim Bilirubin-UGT, tdk seberat
tipe I, bilirubin serum tdk sampai 20 mg/dL
- Dapat di Tx dg Phenobarbital dosisi tinggi
E. Gilbert Syndrome
- Mutasi pada gen pengkode enzim Bilirubin-UGT, sering pada pria
- Klinis tdk berat, aktivitas enzim masih ada sekitar 30 %
F. Toxic Hyperbilirubinemia
- chloroform, arsphenamine, CCl4, acetaminophen, virus hepatitis,
chirrhosis, jamur beracun kerusakan parenkim hati

CONJUGATED HYPERBILIRUBINEMIA
A. OBSTRUKSI SALURAN EMPEDU
- Penyebab: baru empedu, ca. caput pankreas.
- Bilirubin diglukoronida tidak bisa diekskresi regurgitasi ke venavena di liver dan saluran limfe bilirubin masuk ke aliran darah dan
urine (choluric jaundice)
- Cholestatic Jaundice: extrhepatic obstructive jaundice
B. DUBIN-JOHNSON SYNDROME
- Autosomal resesif, mutasi gen MRP-2
- Hepatosit pada area centrilobular mengandung pigmen hitam yang
abnormal yang merupakan derivat epinephrine.
C. ROTOR SYNDROME
- Hiperbilirubinemia terkonjugasi yang kronis, histopatologi hepar tetap
normal
- Etiologi: tdk tahu

Diagrammatic representation of some major causes of jaundice

Prehepatic indicates events in
the blood stream, the major
cause would be various forms
of hemolytic anemia
Hepatic signifies events in the
liver, such as the various types
of hepatitis or other forms of
liver disease (eg. cancer)
Posthepatic refers to evens in
the billiary tree, the major
causes of posthepatic jaundice
are obstruction of the common
bile duct by a gallstone (billiary
calculus) or bya cancer of the
head of pancreas

Laboratory Results in Normal Patients and Patients with Three

Different Causes of Jaundice
Condition

Serum Bilirubin

Urine
Urobilinogen

Urine
Bilirubin

Fecal
Urobilinogen

Normal

Direct: 0.10.4
mg/dL
Indirect: 0.20.7
mg/dL

04 mg/24 h

Absent

40280 mg/24 h

Hemolytic
anemia

Indirect

Increased

Absent

Increased

Hepatitis

Direct and
indirect

Decreased if
microobstruction is
present

Present if
microobstruction
occurs

Decreased

Obstructive
jaundice

Direct

Absent

Present

Trace to absent