RETNO SINTOWATI

Purin, Pirimidin, Asam

nukleat
I. Pendahuluan
II. Basa Purin dan Basa Pirimidin
III. Nukleosida
IV. Nukleotida
V. Asam Nukleat

I. Pendahuluan

Nukleoprotein

protein majemuk
terdapat dalam inti sel
penyusun material inti sel
tersusun atas :

protein: histon / protamin

gugus non protein
semua sel hidup mengandung nukleoprotein

Kromatin
Konstituen penting dalam sel, tersusun atas

nukleoprotein
kelainan pembentukan nukleoprotein
perubahan dlm pertumbuhan /reproduksi sel
sebab kromatin
pembelahan sel

Nukleosida Purin
Adenosin

Guanosin

adenosin deaminase
fosforilase

NH3

purin nukleosida

Inosin

Guanin

purin nukleosida
fosforilase

Hipoxantin
Xantin oksidase

Xantin + O2

asam urat

urin
Xantin oksidase

Penyelamatan Purin
Untuk pembentukan nukleotida bbrp jaringan
manusia
tgt pd purin /purin ribonukleosida eksogen
Ada 2 jalan penyelamatan :
1. Fosforibosilasi purin bebas dgn enzim yang
membutuhkan PRPP sbg donor ribosa fosfat
2. Fosforilasi nukleosida purin pd gugus 5-hidroksi
oleh enzim adenosin kinase
Contoh :
1. Adenin menjadi AMP dg enzim adenin fosforibosa
transferase
2. Hipoxantin dan Guanin menjadi IMP dan GMP
dengan enzim hipoxantin-guanin fosforibosa
transferase

Nukleosida pirimidin
Sitidin

Deoksisitidin

deaminasi

Uridin

deaminasi

Deoksiuridin Deoksitimidin
fosforilse

Urasil dan Timin

degradasi

-alanin + NH4+ +CO2

degradasi

As. -amino isobutirat + NH4+

+CO2
urine
Banyak tdp pd penderita tumor yang dikhemoterapi
( DNA degradasi )

Kepentingan biomedis
kemampuan nukleotid menyerap sinar UV

menjadikan sinar UV sbg unsur

mutagen yg potent
senyawa analog purin & pirimidin
disintesis secara kimiawi digunakan
untuk terapi penyakit kanker
Sel yg akan membelah, selnya
mengalami replikasi
Katabolisme purin xantin asam
urat

II. Basa Purin Dan Basa

Pirimidin
Basa Purin
Basa heterosiklis , molekul planar , aromatik
sebag. besar tdp. dlm sel dlm. bentuk asam

nukleat
disintesis secara de novo, dg sumber :
N1 dari as. Aspartat,
C6 dr CO2 respirasi,

N7 dari glisin,
C8 dari N5,N10-metenilFH4,
N3 dan N9 dari nitrogen amida der. Glutamin dan
C2 dari N-formil -tetrahidroksi folat

Diberi nomor mulai atom N1 kearah kebalikan

jarum jam

Basa Pirimidin
nama lebih panjang dari purin ttp hy

mgd 6 atom yang heterosiklis, sedang

purin
nama pendek mgd 9 atom heterosiklis
Sitosin (S): tdp dlm semua asam nukleat,

kecuali DNA virus ttt

Timin (T) : trtm tdp dlm as nukleat
yg mgd gula deoksiribosa (DNA);
sedikit tdp dlm tRNA
Urasil (U): tdp dalam RNA

Nitrogenous Bases
Planar, aromatic, and heterocyclic
Derived from purine or pyrimidine
Numbering of bases is unprimed

Nucleic Acid Bases

Purines

Pyrimidines

Sugars
Pentoses (5-C sugars)
Numbering of sugars is primed

Sugars
D-Ribose and 2-Deoxyribose

*Lacks a 2-OH group

Nucleosides
Result from linking one of the sugars with a

purine or pyrimidine base through an Nglycosidic linkage

Purines bond to the C1 carbon of the sugar at

their N9 atoms
Pyrimidines bond to the C1 carbon of the sugar
at their N1 atoms

Nucleosides

Phosphate Groups
Mono-, di- or triphosphates
Phosphates can be bonded to either C3 or C5

atoms of the sugar

Nucleotides
Result from linking one or more phosphates

with a nucleoside onto the 5 end of the

molecule through esterification

III. Nukleosida dan nukleotida

Nukleosida:
- tdr dari basa + d-ribosa gula ribosa

terikat
pada N9 basa purin dan pada N1 basa
pirimidin (kecuali U pd C5 ikatan C-C) dan
semuanya mrpk ikatan -N-glikosidik
Nukleosida yang dimanfaatkan untuk
menghambat pertumbuhan sel:
5-fluorourasil
- 5-iodo-2deoksiuridin
6-tioguanin
- 6-merkaptopurin
6-azauridin
- arabinosil sitosin

Nukleotida
Mrpk unit struktural as nukleat, mrpk komponen seluler

yg mudah dpt diketahui krn menyerap kuat sinar UV, der. basa
purin absorbsinya lebih kuat dari der. basa pirimidin
Tersusun: - basa purin/ basa pirimidin
- gula (ribosa atau 2-deoksiribosa),
- asam fosfat

Pengikatan fosfat: RNA pd at C3 dari gulanya

DNA pd at C5 dari gulanya

Basa

Nukleosida

Nukleotida

Singkatan
RNA

Adenin

Adenosin

Asam adenilat

Guanin

Guanosin

Asam guanilat

Sitosin
Urasil
Timin

Sitidin
Uridin
Timidin

Asam Sitidilat
Asam uridilat
Asam timidilat

A
G
C
U
T

DNA
dA
dG
dC
dU
dT

Lanjutan nukleotida
-

bentuk nukleotida lebih mudah larut dalam air dp nukleosida

dan basa bebasnya

Fungsi nukleotida
1. Berperan dlm metabolisme energi
Contoh: ATP - bentuk energi kimia yang diperlukan
untuk sel, dihasilkan dari fosforilasi
oksidatif atau fosforilasi tingkt
substrat
- terlibat dalam kontraksi otot
- transport aktif
- mempertahankan gradien/ion
- membantu sbg donor fosfat untuk
sintesis nukleosida 5-trifosfat
substrat untuk reaksi yg dikatalisis
oleh RNA dan DNA polimerase

Lanjutan fungsi nukleotida

2. Monomer unit asam nukleat: DNA dan RNA
3. Mediator fisiologik: nukleosida dan nukleotida
membantu sbg kunci mediator fisiologik proses
metabolik:
- adenosin penting dlm kontrol aliran darah koroner
- ADP critical dlm agregasi platelet dan koagulasi
darah
- cAMP & cGMP: bekerja sbg second messenger
- GTP: diperlukan untuk capping mRNA
4. Precursor function: GTP adalah prekursor pembentukan kofaktor, tetrahidrobiopterin, untuk reaksi
hidroksilasi dan generasi oksida nitrat
5. Komponen koenzim: NAD, NADP, FAD

Lanjutan fungsi nukleotida

Activated intermediate
- nukleotida membantu sbg kunci activated
intermediate, diperlukan untuk berbagai reaksi
- UDP-glukosa-kinase intermidiate sintesis
glikogen, glikoprotein dll.
7. Allosteric effectors pd metabolisme nukleotida
6.

Sintesis nukleotida
- Di dalam sel mamalia disintesis secara de novo
- Tidak semua sel mampu mensintesis nukleotid

purin
Contoh; sel-sel darah merah
- Pada akhir reaksi menghasilkan ITP prekursor
adenosin 5-monofosfat (AMP) dan guanosin 5mono- Fosfat (GMP)

Lanjutan sintesis nukleotida

- IMP AMP energinya dari GTP
- IMP GMP energinya dari ATP
- Apabila terjadi defek dlm reaksi metabolik

menyebabkan kehilangan pengaturan

sintesis nukleotida purin over produksi
purin asam urat meningkat
Artritis gout

Pembentukan deoksi ribonukleotida

pada keadaan sel tidak proliferasi

rendah
- pada saat replikasi sintesis DNA sintesis
deoksiribonukleotida meningkat untuk
mensuport sintesis DNA
-

IV. Asam nukleat

(Polinukleotid)
dibentuk dari ikatan fosfodiester yg

terikat pada C3- dan C5 monomer yg

berdekatan
ada mol. yg berakhir 5 dan 3
masing-masing mol. memp. struktur
primernya, contoh 5-3p Gp Gp Ap Tp
Cp A
Ada 2 macam asam nukleat besar: DNA
& RNA

Nucleotide
Metabolism
PURINE RIBONUCLEOTIDES: formed de novo
i.e., purines are not initially synthesized as free bases
First purine derivative formed is Inosine Mono-

phosphate (IMP)

The purine base is hypoxanthine

AMP and GMP are formed from IMP

Purine Nucleotides
Get broken down into Uric Acid (a purine)

Buchanan (mid 1900s) showed where

purine ring components came from:
N1: Aspartate Amine
C2, C8: Formate
N3, N9: Glutamine
C4, C5, N7: Glycine
C6: Bicarbonate Ion

CO2
Aspartate

Formyl-THF

Glycine

H
C
HN

HC

Glutamine

N
H

H
N
C
N
H

Formyl-THF

Glutamine

H
C
HN

HC

H
N
CH

N
H

N
H

Glycine
Glutamine (2)
Aspartate
N10-Formyl-THF (2)
CO2

H
C
HN

CH

HC

CH
N
H

Purine

Pyrimidine

(C5H7N4)

(C4H6N2)

Glutamine
Aspartate
CO2

Purine Nucleotide Synthesis

OOC

2-

O3P O CH2

H
OH

OH

OH

O3P O CH2

ADP
+ Pi

C
HC

N
H

CH2

N
SAICAR Synthetase

AIR
Car boxylase

ADP + Pi

COO

H
OH

Ribose-5-Phosphate

Fumarate

5-Aminoimidazole Ribotide (AIR)

ADP + Pi

AIR
Synthetase

Glutamine
+ H2O
Amidophosphoribosyl
Transferase

NH2

OH

OH

HN

Formylglycinamidine ribotide (FGAM)

ADP
+ Pi

H2C

NH2

NH

H2 C
C
N10-Formyl-THF

THF

Glycinamide Ribotide (GAR)

GAR Transformylase

C
H

CH

NH

Ribose-5-Phosphate

5-Formaminoimidazole-4-carboxamide
ribotide (FAICAR)

ATP +
Glutamine +
H2O

IMP
Cyclohydrolase

O
C

CH

HN

HC

C5

NH

H
OH

H2O

H
N

H
OH

ADP +
Glutamate + Pi

FGAM
Synthetase

GAR Synthetase

Ribose-5-Phosphate

C4

NH

Ribose-5-Phosphate
Formylglycinamide ribotide (FGAR)

AICAR
Transformylase

THF

C
H2N

Glycine
+ ATP

CH

-5-Phosphoribosylamine (PRA)

O3P O CH2

N10-FormylTHF

2-

5-Aminoimidazole-4-carboxamide
ribotide (AICAR)

H
N
H2C

O3P O CH2

Ribose-5-Phosphate

ATP

Glutamate
+ PPi
2-

CH

H2N

Ribose-5-Phosphate
5-Phosphoribosyl--pyrophosphate (PRPP)

C4

H2N

Adenylosuccinate
Lyase

H2N

CH

5-Aminoimidazole-4-(N-succinylocarboxamide)
ribotide (SAICAR)

HC 4

CH

H2N

ATP
+HCO3

C4

Ribose-5-Phosphate

Ribose
Phosphate
Pyrophosphokinase

H
OH

Carboxyamidoimidazole Ribotide (CAIR)

AMP

2-

Aspartate
+ ATP

CH

H2N

-D-Ribose-5-Phosphate (R5P)
ATP

C4

O
COO

CH
N

N
2-

O3P O CH2
H
H
OH

H
H
OH

Inosine Monophosphate (IMP)

IMP is converted to AMP or GMP

GTP
Aspartate

O
HN
HC

C
C

GDP + Pi
Fumarate

NH2
N

CH

HC

CH

Ribose-P
H2O + NAD+

Ribose-P

IMP

AMP

NADH + H+
O
HN
HO

C
C

ATP
Glutamine

ADP + Pi
Glutamate

HN

CH
H2N

C
C

CH
N
Ribose-P

Ribose-P

XMP

GMP

Regulatory Control of Purine

Nucleotide Biosynthesis
GTP is involved in AMP synthesis and ATP is involved

in GMP synthesis (reciprocal control of production)

PRPP is a biosynthetically central molecule (why?)
ADP/GDP levels negative feedback on Ribose Phosphate

Pyrophosphokinase
Amidophosphoribosyl transferase is activated by PRPP levels
APRT activity has negative feedback at two sites

ATP, ADP, AMP bound at one site

GTP,GDP AND GMP bound at the other site

Rate of AMP production increases with increasing

concentrations of GTP; rate of GMP production

increases with increasing concentrations of ATP

Regulatory Control of Purine

Biosynthesis
At level of IMP production:
Independent control
Synergistic control
Feedforward activation by PRPP

Below level of IMP production

Reciprocal control

Total amounts of purine nucleotides controlled

Relative amounts of ATP, GTP controlled

Purine Catabolism and Salvage

All purine degradation leads to uric acid (but it might

not stop there)

Ingested nucleic acids are degraded by pancreatic
nucleases, and intestinal phosphodiesterases in the
intestine
Group-specific nucleotidases and non-specific
phosphatases degrade nucleotides into nucleosides
Direct absorption of nucleosides
Further degradation

Nucleoside + H2O base + ribose (nucleosidase)

Nucleoside + Pi base + r-1-phosphate (n. phosphorylase)
NOTE: MOST INGESTED NUCLEIC ACIDS ARE DEGRADED AND
EXCRETED.

Intracellular Purine Catabolism

Nucleotides broken into nucleosides by action

of 5-nucleotidase (hydrolysis reactions)

Purine nucleoside phosphorylase (PNP)
Inosine Hypoxanthine
Xanthosine Xanthine
Guanosine Guanine
Ribose-1-phosphate splits of
Can be isomerized to ribose-5-phosphate

Adenosine is deaminated to Inosine (ADA :

Adeosine deaminase)

Intracellular Purine Catabolism

Xanthine is the point of convergence for

the metabolism of the purine bases

Xanthine Uric acid
Xanthine oxidase catalyzes two reactions

Purine ribonucleotide degradation pathway is

same for purine deoxyribonucleotides

Adenosine Degradation

Xanthosine Degradation

Ribose sugar gets recycled (Ribose-1-Phosphate R-5-P )

can be incorporated into PRPP (efficiency)
Hypoxanthine is converted to Xanthine by Xanthine Oxidase
Guanine is converted to Xanthine by Guanine Deaminase
Xanthine gets converted to Uric Acid by Xanthine Oxidase

GMP

Guanosin

AMP

IMP

Guanin

Inosin

Hipoxantin

Xantin

Asam urat URIN

tmp kerja xantin oxidase, dihambat oleh Alopurinol

Uric Acid Excretion

Humans excreted into urine as insoluble crystals
Birds, terrestrial reptiles, some insects excrete

insoluble crystals in paste form (hewan urikotelik)

Excess amino N converted to uric acid

Others further modification :

Uric Acid Allantoin Allantoic Acid Urea Ammonia
Ekskresi asam urat total man 400-600mg/24jam
Aspirin dosis tinggi hambat ekskresi & reabsorpsi urat.

Purine Salvage
(Jalur penyelamatan purin)
Adenine phosphoribosyl transferase (APRT)

Adenine + PRPP AMP + PPi

Hypoxanthine-Guanine phosphoribosyl

transferase (HGPRT)

Hypoxanthine + PRPP IMP + PPi

Guanine + PRPP GMP + PPi
(NOTE: THESE ARE ALL REVERSIBLE REACTIONS)

AMP,IMP,GMP do not need to be resynthesized

de novo !

Di hati : nukleotida defosforilasi

nukleosida
Sering nukleosida diputus basa bebas +
gula
nukleosida & basa dibawa ke jar lain

refosforilasi mjd nukleotida

Basa bebas diselamatkan dg mereaksikan dg
PRPP nukleotida baru
Enzim penghemat basa tsb HGPRT
Reaksi analog juga terjadi utk pirimidin

A CASE STUDY : GOUT

A 45 YEAR OLD MAN AWOKE FROM SLEEP WITH A PAINFUL

AND SWOLLEN RIGHT GREAT TOE. ON THE PREVIOUS NIGHT

HE HAD EATEN A MEAL OF FRIED LIVER AND ONIONS, AFTER
WHICH HE MET WITH HIS POKER GROUP AND DRANK A
NUMBER OF BEERS.
HE SAW HIS DOCTOR THAT MORNING, GOUTY ARTHRITIS
WAS DIAGNOSED, AND SOME TESTS WERE ORDERED. HIS
SERUM URIC ACID LEVEL WAS ELEVATED AT 8.0 mg/dL (NL <
7.0 mg/dL).
THE MAN RECALLED THAT HIS FATHER AND HIS
GRANDFATHER, BOTH OF WHOM WERE ALCOHOLICS, OFTEN
COMPLAINED OF JOINT PAIN AND SWELLING IN THEIR FEET.

A CASE STUDY : GOUT

THE DOCTOR RECOMMENDED THAT THE MAN USE

NSAIDS FOR PAIN AND SWELLING, INCREASE HIS

FLUID INTAKE (BUT NOT WITH ALCOHOL) AND
REST AND ELEVATE HIS FOOT. HE ALSO
PRESCRIBED ALLOPURINOL.
A FEW DAYS LATER THE CONDITION HAD
RESOLVED AND ALLOPURINOL HAD BEEN
STOPPED. A REPEAT URIC ACID LEVEL WAS
OBTAINED (7.1 mg/dL). THE DOCTOR GAVE THE
MAN SOME ADVICE REGARDING LIFE STYLE
CHANGES.

Impaired excretion or overproduction

of uric acid
Uric acid crystals precipitate into joints
(Gouty Arthritis), kidneys, ureters
(stones)
Lead impairs uric acid excretion lead
poisoning from pewter drinking goblets
Fall of Roman Empire?

Xanthine oxidase inhibitors inhibit

production of uric acid, and treat gout

Allopurinol treatment hypoxanthine
analog that binds to Xanthine Oxidase
to decrease uric acid production

ALLOPURINOL IS A XANTHINE OXIDASE

INHIBITOR
A SUBSTRATE ANALOG IS CONVERTED TO AN
INHIBITOR, IN THIS CASE A SUICIDE-INHIBITOR

Pyrimidine Ribonucleotide
Synthesis
Uridine Monophosphate (UMP) is synthesized

first
CTP is synthesized from UMP

Pyrimidine ring synthesis completed first;

then attached to ribose-5-phosphate

N1, C4, C5, C6 : Aspartate

C2 : HCO3N3 : Glutamine amide Nitrogen

Pyrimidine Synthesis

2 ATP + HCO3- + Glutamine + H2O

2 ADP +
Glutamate +
Pi

Carbamoyl
Phosphate
Synthetase II

NH2

CH

C
N
H

PO3-2

PRPP

PPi

2-

O3P

Orotate Phosphoribosyl
Transferase

Pi

HO

N
H

H2O

Dihydroorotase

Orotidine-5'-monophosphate
(OMP)

OMP
Decarboxylase
CO2

C
CH

HN
C

N
H
Dihydroorotate

COO

O3P

CH2

CH
N

O
2-

CH

COO

Carbamoyl Aspartate

OH

OH

CH2

HN
C

CH

Quinone

CH2

COO

Dihydroorotate
Dehydrogenase

NH2

Reduced
Quinone

Orotate

Aspartate
Transcarbamoylase
(ATCase)

CH2

COO

Carbamoyl Phosphate
Aspartate

HN

CH

HN

OH

OH

Uridine Monophosphate
(UMP)

UMP UTP and CTP

Nucleoside monophosphate kinase

catalyzes transfer of Pi to UMP to form UDP;

nucleoside diphosphate kinase catalyzes
transfer of Pi from ATP to UDP to form UTP
CTP formed from UTP via CTP Synthetase

driven by ATP hydrolysis

Glutamine provides amide nitrogen for C 4 in

animals

Regulatory Control of
Pyrimidine
Synthesis
Animals regulation
at carbamoyl phosphate
synthetase II
UDP and UTP inhibit enzyme; ATP and PRPP

activate it
UMP and CMP competitively inhibit OMP
Decarboxylase

*Purine synthesis inhibited by ADP and GDP at

ribose phosphate pyrophosphokinase step,
controlling level of PRPP also regulates
pyrimidines

Caused by defect in protein chain with

enzyme activities of last two steps of

pyrimidine synthesis
Increased excretion of orotic acid in
urine
Symptoms: retarded growth; severe
anemia
Only known inherited defect in this
pathway (all others would be lethal to
fetus)
Treat with uridine/cytidine
HOW DOES URIDINE AND CYTIDINE

ADMINISTRATION WORK TO TREAT OROTIC

ACIDURIA?

Degradation of
Pyrimidines
CMP and UMP degraded to bases similarly to
purines

Dephosphorylation
Deamination
Glycosidic bond cleavage

Uracil reduced in liver, forming -alanine

Converted to malonyl-CoA fatty acid
synthesis for energy metabolism

Deoxyribonucleotide
Formation
Purine/Pyrimidine degradation are the same
for ribonucleotides and deoxyribonucleotides
Biosynthetic pathways are only for
ribonucleotide production
Deoxyribonucleotides are synthesized from
corresponding ribonucleotides
Pd tingkat difosfat,gugus ribosa direduksi mjd

deoksiribosa o/ enz Ribonukleotida reduktase.

ex. CTP defosforilasi CDP reduksi dCDP
dCTP defosforilasi dan deaminasi dUMP

Katabolisme pirimidin menghasilkan metabolit

yg dpt larut dalam air, yaitu : CO2, NH3, Balanin,

B-aminoisobutirat
Produk katab purin tidak begitu mudah larut,
kelarutan ditingkatkan dg alkalinisasi.