PRPP

Anabolam SAICAR
A Bummary o
Catabolam

T savaga
AICAR
10 biosynthotic
s0ps Metabolism
FAICAR

of
AMP S-AMP MP XMP GMP

Adenosin Inosino
XanthosinaGuanosine
Nucleotides
I
A ypoxanithino Guanino

Xanthino

Agurtal
Uric acid

hals
Cemeric Pyrd bese by
Agr RAMON S. DEL FIERRO, Ph.D. (Tol
ormte
Farmn Professor of Biochemistry
Atthe end of the Session, thestudents shoula oea
answer the following

1. Identify the "De Novo'" and the "Salvage Pathway o

purine and pyrimidine nucleotides putting emphasis
on:
1.1 Regulated and/or rate limiting enzymes and
reactions.
1.2 Positive and negative modulators that affect the

rate of synthesis.
1.3 Substances that serve as sources of the carbon
and nitrogen atoms of the purine and pyrimidine
rings.
1.4 Difference between the de novo synthesis of
purine and pyrimidine nucleotides.
Attheend of the Session,the students should beabe to
answer the following

2. are purine and pyrimidine nucleotides

How
What is the clinical significance of the
degraded?
end products especially uric acid?

3. What are the biochemical basis of some inborn

errors of metabolism?
3.1 Gout
3.2 Lesch-Nyhan syndrome
3.3 Adenosine Deaminase (ADA) Deficiency
3.4 Purine nucleoside phosphorylase deficiency
3.5 Orotic aciduria
 New enzyme
Enryma Adenine
intermediates

OH OH

Novel
cofactors

R1 subuni

OO-0-

Currentlyused
NDP
phototherapy treatment fpr
Peductene
2 subunis

hyperbilirubinemia
Radical reaction mechanisms
62025
 Biosynthesis of Nucleotides
De novo Biosynthesis
refers the synthesis of complex molecules from
to
or amino acids,
simple molecules such as sugars
as opposed to recycling after partial degradation.

Salvage Pathways
Are pathways in which nucleotides are synthesized
from intermediates in the degradative pathway for
nucleotides.

They are used to recover bases and nucleosides

that are formed during degradation of RNA and
DNA
 H
N7
N1 8CH
2 9
HC3
N1 -N
H
Purine
 CO2
Glycine
Aspartate

HN
CHO

Formate
9coM
co
NN 1
Formate
N0-Formyl THF

NoTFormyl THF
Amide N
ofglutamine
 HaN H-COH
H
CO2
Glycine
Aspartate

CHO
9coM
co
N N Formate
N,

N CHO
N:0-FormylTHF
çoM

Formate
Nio-Formyl THF
HNNN
Amide N NIl

ofglutamine 2--CI--coo
 Olamine phonphonibonylis GeingA1 5Glycinderibonscleotda
NlO

NFormylcycinamide ribonoclaoti

Pentose Phosphate
Pathway &ATP

N-Formylycinemidine ribanuclet

ATP

S-Aminoimidarole nibonucleotide

RIboso 1-phoaphets HCO

Inosine 5-monophosphat
SAminoimidaolecarbanylat
nbonucleotide

ATP&
Aupartate

5-Formamidoimidazolon 5-Aminoimidaolo4 S-Aminoimidanolo-4-N-euocinocarboam

crbonmide riboeucleotida carboamida nboeacleotida
rbacleoeid
2021
116
 Purine Synthesis

Purine synthesis is critical to fetal development,

therefore defects in enzymes wll result in a nonviable
fetus.

PRPP synthetase defects are known and have severe

consequences
PRPP synthetase superactivity has been dccumented,
resulting in increased PRPP, elevated
levels of
nucleotides, and increased excretion of uric acid.

16202
 O-GH2 O H From: Pentose
Phosphate

o-- Pathway

5-Phosphoribosyl
OH H 1-pyrophosphate (PRPP
NI
Glutamine 1-c-cil-Ci-c-coo
glutamine-PRPP
amidotransferase Glutamate
PPi
No
-0-CH2 NH2
H 5-Phospho-p-
D-ribosylamine
OH OH
162023
 o-CH2 0 NH2
H5-Phospho-B-
D-ribosylamine
OH O
-Glycine HN-CH--OH

GAR synthetase ATP

ADP+P
CsNH3
H2 Glycinamide
O=C No
O ribonucleotide (GAR)
C NH
16
3023 R
 NH3 Glycinamide
H2C ribonucleotide (GAR)
O=C o
NH CHO

R
N0-Formyl H, folate
GARtransformylase
Ha folate
H
Cs
H2 G-H Formylglycinamide
O=C ribonucleotide (FGAR)
 H2 Formylglycinamidine
HN- NH ribonucleotide (FGAM)

FGAM Cyclase ATP

LAIRsynthetase) ADP+ Pi
H20
HC CH 5-Aminoimidazole
CH
ribonucleotide (AIR)
H2N
 CH -Aminoimidazole
ribonucleotide (AIR)
H2N
HCO
AIR N-CAIR ATP
carboxylase synthetase
ADP Pi
CO2 HC CH N3-Carboxyaminoimidazole
ribonucleotide
(NS-CAIR

NS-CAIR
mutase
ooCC6
CH
H2N N Carboxyamino-
imidazole ribonucleotide
(CAIR)
 ooc
Carboxyamino
imidazole ribonucleotide
HaN (CAIR)

Aspartate

SAICAR synthetase
ATP
ADP+Pi
coo

CH N-Succinyl-5-aminoimidazole-4-
coo carboxamide ribonucleotide (SAICAR)
H2N
 N
COO
H2

coo
H2N
CH
N
N-Succinyl-5-aminoimidazole-4-
carboxamide ribonucleotide (SAICAR)

H CooH

SAICAR Iyase Fumarate

C
HOOC

H2N CH 5-Aminoimidazole-4-carboxamide
ribonucleotide (AICAR)
H2N
 H2N CH 5-Aminoimidazole-4-carboxamica
ribonucleotide (AICAR)
HN
R
CHO N co
çoH

AICAR N10-Formyl Hafolate

transformylase
Hafolate

H2N CH N-Formylaminoimidazole
4-carboxamide
C2 ribonucleotide (FAICAR)
O-CN
HH
 H2N CH N-Formylaminoimidazole-
4-carboxamide ribonucleotide
(FAICAR)
R
IMP synthaseH20

HN CH
HCN
o-CH2 Inosinate (IMP)
O

OH OH
 Purine Synthesis
PAP?alamins Sphong honbonmylamina GlycinsA sGlycinmde nibonucleotid

NFormylsheinnide ribonclectid

dlutamina
&ATP

NFormylycinamidine rnibomuclectida

ATP

S-Aminoimidazola nibonuclectide

Fiboeo 1-phosphato HCO

Inosine S-moophosphato
S-Amincimidazolecarbonylata
nboeucleotido

ATP&
Burnarnta Aupartate

SFormamidoimidazolo-Ao S-Aminoimidaolo-4 S-Aminoimidanole 4-N-oucinocarborama

carboamida ribonuclectida cebeamide nbooucleotida
 From the Preformed PRPP
1. Ng from the amide group of Glutamine

2. Ca, Cs, N,from Glycine

3. Cg from Nio-formyl tetrahydrofolic acid

4. Ng from the amide of Glutamine

5. C from CO2
6. N, from Aspartic Acid

7. Ca from Ni0-formyl tetrahydrofolic acid

1/162023
blaaShahm dU7)
ashahas
Lnite (CAI)
Cahaqandamtna
De Novo
Biosynthesis

KaeuSPhaphas Tbedheyhan

dcaaa
rbetta (AICA

O
O=CN
Rbeu-Pohatn
B3aghribaylame G1A
Fermyghyeinai hendeFOAMD Sharamiimidle4aeumite
etd(AICAS

Ca
0-0-
TAeS-Pephan
ypoxannine-GtanineT0SprT*
Transferase Activity
PRPP

De novo synthesis

P AMP
PRPP

Guanosin Adenosine
Inosinet APRT
Salvage
HGPRT
PNP
Guanine Hypoxanthino Adenino
PRPP

Xanthine

xo Catabolism

acld
Uric

Unicase

Allantoln
 H

N3 5CH
6CH
HC
Pyrimidine

1/16/2023
BTosynthesis of AMP and GMP from
IM
Hadacidin
(N-formy-N- 0OC-CH--cOO
hydroxyglycine) is an Fumarate
NH NHa
analog of L-aspartate

GTP GDP+P
adenylosuccinat
Aspartate lyase

adenylosuccinate
RID-
Adenylosucinate Adenylate
synthetase
(AMP)

RIB-
H,0
Inosinate
NAD
(IMP)
NADH+H
Gln Glu ATP AMP PP
IMP
debydrogenase
HN
XMP-glutamine
H0 amidotransferase
Mycophenolic acid (MPA) RIb-
Xanthylate Guanylate
(XMP) (GMP)
 Wcarbamoylaspartat
diydreeretase
2coo
LOIydroorotate
n-coo
NAD
egenasa NADH
Oretat
PAPP
oo
oretate
phespheribesyi-

Biosynthesis of
anateate PP
Orotltytate
O-o
orotidylate Pyrimidine Nucleotides
decarbaxylase
-co
Urldylate
(UMP) O-o- from Orotidylate
kinases
.26
2ADP
Urldlne 5triphosphate (UTP)
GIn
alu
idylate
ynthetase

ADP P

o0-0-
-yidine steriphosphate (CTP),
 Lesch-Nyhan Syndrome
Guanine Phosphoribosyl Transferase
Hypoxanthine
(HGPRTase) deficiency
Gene;
X-linked genetic condition encoded by the HPRT1
in the gene is carried by the mother
present baby boys;
and passed on to her son

not used the salvage

Hypoxanthine and Guanine are in
nucleotide biosynthesis
pathways of purine

and Guanine are not utilized repeatedly but

Hypoxanthine
converted into uric acid

Severe neurologic disease, characterized by self-

mutilating behaviors
such as lip and finger biting and/or
head banging
 Pyrimidine Ribonucleotide Synthesis

O Uridine Monophosphate (UMP)is synthesized ffrst

from UMP
CTP is synthesized
first; then attached to
O. Pyrimidine ring synthesis completed
ribose-5-phosphate

3N 5 Ni, Ca, Co, Co: Aspartate

C2:HCO3
Ng: Glutamine amide Nitrogen
6

1/162023
iypoxannine-GUaNHAe FHOspTo
Transferase Activity
De Novo Synthesis

Ribose-5-P

PRPP
5-Phosphoribosyl.
Salvage Synthesis glycinamide
GMP IMP

Guanosine Inosine
HPRT

Guanine Hypoxanthine

Deoxyinosine
5-Amino-4-Imidazole Xanthine
Carboxylate
URIC ACID
Formiminoglycine

116/2023
 Pyrimidine Synthesis
is critical to fetal development just-as
Pyrimidine Synthesis
is critical.Therefore an absolute
purine metabolism
of pyrimidine synthesis would
deficiency of an enzyme
be fatal.

A very low level of the enzyme UMP synthase has been

documented, resulting in the condition orotic aciduria.

1162023
 phosphate
aspartate
trans
carbamoylase P HN
N-Carbamoylaspartate -COO
dihydroorotase

H0
L-Dihydroorotate HN
H-Co0
NAD O
dihydroorotate
dehydrogenase

NADHH
Orotate

PRPP
orotate
phosphoribosyl-
transferase
PP
Coo
Orotidylate -0-CH

OH OH
 Coo
Orotidylate -0-H
orotidylate
HN
decarboxylase
c02
(UMP)
Uridylate -0-gH

kinases
2T
2 ADP
Uridine 5-triphosphate (UTP)

-Gln
Glu
cytidylate
synthetase
ATP NH
ADP Pi

---0-ta
----- cytidine 54triphosphate (CTP),N
1162023
 Synthesis of CTP and TMP from UMP
UMP

ACP NADPH H NADP

UDP dUDP(dayurisino diphoephate)

nubOMUCLEOTDC
HO
ATP REDUCTASE

ACP
dUMP
P
UTP

ATP N'NMethoteoa l,folete

alutarine THYLIDYLATE
CTP OYTUcC
THAE H, tolato

CH3

A-s4-- ah-s-
CTP TMP

162023
 Orotic Aciduria
Caused by defect in protein.chain with
enzyme activities of tast two step5 9f
Pyrimidine synthesis
a
Typel-reflects deficiency of both
orotate phosphoribosyl transferase and
orotidylate decarboxylase

Clinical Type Il-is due to deficiency only of

orotidyate decarboxylase
CoTelation
Increased excretion of orotic acid in urine:
Pyrimidine synthesis is decreased

Symptoms: retarded growth; severe anemia

Only known inherited defect in this

pathway (all others would be lethal to
fetus)
1/16/2023
 dNDP NDP
HO

Ribonucleotides
areprecursors
Ribonucleotide Ribonucleotide of Deoxyribo-
reductase reductase
HS
Nucleotides

SH
Glutaredoxin) Glutaredoxin Thioredoxin) Thloredoxin
SH sH
E.coli
glutaredoxin
reductase
thioredoxin
reductase
Ribonucleotide
GSSG FAD
2GSH FAPH Reductase
glutathlone
reductase
NADPH+H* NADP NADPH+H NADP
 ThioredOxin-
Physiologic reducing agent of RNR
Cys pair can swap H atoms with disulfide formed
regenerate original enzyme
Thioredoxin gets oxidized to disulfide
SE

Reduced Thioredozin Oidized Thioredoxin

Oxidized Thioredoxin gets reduced by NADPH(final electron acceptor)

1162023
 Thymidylate isderived from
dCDP and dUMP
CDP dCDP nucleoside
dCTP
ribonucleotide deaminase
diphosphate
reductase kinase
cinase dUTP
UDP dUDP.
dUTPase

dUMP
thymidylate
synthase

dTMP
1/16/2023
 Purine Degradation

O Purine Nucleotides from ingested nucleic acids or

turnover of cellular nucleic acids is excreted by
humans as Uric Acid.

O Humans excrete about 0.6 g uric acid every 24 hours.

116/2023
 Glutamine

via Carbamoyl
N Aspartate
phosphate
CO,
Ribose 5-phosphate

Generic pyrimidine base

116/2023
 Purine Catabolism and Salvage
o Allpurine degradation
leads to
UricAcid (but
it might not
stop there)
O Ingested Nucleic Acids are degraded
to Nucleotides by
Pancreatic Nucleases, and Intestinal Phosphodiesteras
es
in the Intestine

16 2023
 Purineand Pyrimidine Salvage
O Free Purine and Pyrimidine bases are constantly
released in cells during the metabolic degradation ui
nucleotides.

O Free Purine and Pyrimidine bases are in large part

salvaged and reused to make nucleotides.

much
Salvage of free nucleotides consumes
less
O
and is the
energy than de novo nucleotide synthesis
preferred source of nucleotides for
energetically
nucleic acid synthesis.

50
116/2023
 dUMP dTMP

O-- o -o-4

thymidylate
synthase Conversion of
dUMP to dTMP
HN by Thymidylate
HN
HN-R Synthase and
N"N-Methyl ene 7,8-Dihydrofolate Dihydrofolate
tetrahydrofolate

Glycine NADPH H*.Synthase

er dihydrofolate
PLP ethy
hydrerymethy
transferase
Serine NADP

Tetrahydrafolate
HNR
 Lesch-NyhanSyinToiroiTG
the urine than normal
Up to 20 times the uric acid in in

individuals. Uric acid crystals form ir the urine.

Treated with allopurinol,

a competitive inhibitor of
xanthine Oxidase.

Symptoms
Severe Gout
Severe mental and physical problems

116/2023
2
ATP HCO,-Glutamine H,O

Srau
HN 0-0CH
Orotate Phopbonborni1
NH:
Transferae

0-C H Coo
Oroti dine-$-monop bosph
0-PO Orotate (ONP)

Carbamoyl Phosphate
Reduced
ONP
QaineneDubydroa Decarbarytasn
ApaoApar otu

O
Trcabyla3e Deydrogma
ATCe) Quinane

HO- CH
CH
H
Diydroorocas
O H coo
Coo
Dihydroorotate
Uridine
Carbamoy1 Aspartate Menop hosphate
(UMP)
 Thymidylate is derived from
dCDP and dUMP
CDP dCDP dCTP
nucleoside
ribonucleotide deaminase
diphosphate
reductase
kinase
UDP dUDP dUTP
dUTPase

dUMP
thymidylate
synthase

dTMP
DESTINY OFNITROGENOUSBASESFENT
AND PHoSPHORIC ACIDS IN THE ORGANISM

Nitrogenous Oxidation to End Products

the
bases

Oxidation with energy formation;

Pentoses synthesis of Nucleotides; synthesis of

Hexoses; synthesis of Coenzymes

Phosphorylation,
ATP synthesis;
of Phospholipids;
synthesis
Phosphoric acid Buffer systems;
constituentof bones, cartilages
 ENNTESTNE

Nucleoproteins (nucleic acids+ proteins)

Nucleic acidsHistones, protamines

Oligonucleotides
Phonoulestor
Mononucleotides
-Phosphanses

Nucleosides+Phosphoric acid
-
Nucloosldases

Nitrogenous
bases
 Intracellular Purine Catabolism

Nucleotides broken into nucleosides by action of

5-nucleotidase (hydrolysis reactions)

Purine Nucleoside Phosphorylase (PNP)

Inosine Hypoxanthine
XanthosineXanthine
GuanosineGuanine
Ribose-1-phosphate splits off

Adenosine is deaminated to Inosine (ADA)

1/16/2023
 Intracellular Purine Catabolism

Xanthine is the point of convergence for the metabolism

of the purine basess

Xanthine Uric acid

Xanthine oxidase catalyzes two reactions

1/16/2023
 Intracellular Purine Catabolism

Nucleotides broken into nucleosides by action of

5"-nucleotidase (hydrolysis reactions)

Purine Nucleoside Phosphorylase (PNP)

InosineHypoxanthine
XanthosineXanthine
GuanosineGuanine
Ribose-1-phosphate splits off

2023
1/16
 GMP

P NHs
Guanosine
P
d
ANP

Adenosine
H20
Inosine
co
c-OH
c-OH
a
Uric acld
Excreted by
Primates, blrda.
reptiles,Insects

osidas Allantoln Most mammal

Ribese Ribose

Hypoxanthin
(keto form
H,O
lamtolnase

Guanin
Hao
enidase Allastoate Bony fishes

Xanthine
tketo form) aengolcase
coo
CHO
h H.O Glyoxylate

Amphiblan
2MNN Urea cartilaginous
fishes

urease
2H0
2co
 Purine Salvage

Salvage of the free purine bases guanine and

hypoxanthine (the deamination product of adenine)
often involves the enzyme Hypoxanthine-Guanine
PhosphoRibosylTransferase (HGPRT)

Salvage of free adenine is accomplished by the

enzyme Adenine PhosphoRibosyl Transferase
(APRT),converting free Adenine and PRPP to
Adenosine MonoPhosphate (AMP)

116/2023
 Xanthine Oxidase

A homodimeric protelin

Contains electron transfer proteins

FAD
Mo-pterincomplex in +4 or +6 state
Two 2Fe-2S clusters
 I1,0

GMP
-aucleotidan

Adenoan
Purine
11,0
penosine
Bemuclestidase -P deaminase NH Degradation
Guanosine Inosine

1I1,0
nuclem nucleosidase
Rbose Ribose

IIN IN IHypoxanthine
(ceto form)
11NN Guanine

thine
oxidase
1,0 O
1,0

deamina
IN Xanthine

HON Cketo
form

ne O+0,
axidase11,0

IN
-oH
 OH OH

CH
HC HC
N
Allopurinol Hypoxanthine
(enol form)
xanthine
oxidase
Excess Uric Acid
OH causes Gout, treated
with Allopurinol,
inhibitor of Xanthine
Oxidase
HO
Oxypurinol
 Kidney Stones

When uric acid is present in high

concentration in the blood, it
may
precipitate as a salt in the kidneys.

The salt can form stones, which can,

in tun cause pain, infection and

kidney damage.
Uric Acid
Crystals
In Big
Toe Joint
 Allopurinol and Hypoxanthine
OH
HN HN Hypoxanth
keto
forn
HN N
HC Guanine
H,O xanthineH,0
0,
uxidan

NH
Allopurino
guanine

Structural Analogs
deaminnse IN Xanthine
Cketo form}

OH HO
xanthineH0+0
oxidaseH,O
CH
HC
Hypoxanthine
HN
HO
o
Urie cid
(enol form)
 Gout
Uric acid crystals will form in the
extremities with a surrounding
area of inflammation. This is
cailed a tophus and is often

described
as an arthritic "great
toe"
Can be caused by a defect in an
enzyme of purine metabolism or
by reduced secretion of uric acid
into the urinary tract.

tophus
Uric Acid Crystals
 Adenosine Deaminase (ADA)Deficiency

A deficiency of ADA causes a moderate to complete lack of

immune function.

It is an immunodeficiency disease in which

associated with
both thymus-derived lymphocytes (B cells) are sparse and
dysfunctional.

Affected children cannot survive outside a sterile environment.

 GOUT
Gout is caused by
precipitation of sodium urate
crystals in the joints resulting
in inflammation ahdpain
UricAcld
Cyatas
ToeJolnt

-
 Uric Acid Excretion

Humans- excreted into urine as insoluble

crystals

Birds, terrestrial reptiles, some insects

Excess amino N converted to uric acid

Others-further modification:
Uric Acid Allantoin Allantoic AcidUrea
Ammonia
Purine Nucleoside Phosphorylase(NP)
Deficiency

Purine Nucleoside Phosphorylase Deficiency is associatd with

severe deficiency of T-cells but apparently normal B
ceilfunction.

Immune dysfunctions appear to result from accumulation of

dGTP and dATP, which inhibit ribonucleotide reductase and
thereby delete cells of DNA precursors.
 -a suicide inhibitor used to treat Gout
Allopurinol

HN
HN
HO HCS

Hypoxanth ine Allopurina

Guanine
Xanthine oxidase

Xanthine Urate
Xanthine
oxidase
Hypoxanthine
d by allopurinol
Inhibite
 Hyperuricemia
urate (üric acid) level
Characterized by plasma
greater than 7.0 mg/dL

Normal plasma levels

Females 2.4-6mg/dL
Males 3-4-7mg/dL

Primary Hyperuricemia
-
an innate defect in
purine metabolism and/or uric acid excretion
 Thmina

Gahydregae
NADP

o, Dihytrothymine

Catabolism
diydropyrms

Of
p-Ureidoisobutyrate
CH
-H,o
pureprepion Pyrimidines
NH HCo
-o
HN-CH9 p-Aminolsobutyrate

Ketoglutarate

Glutamata

Mathylalomy
dl
Deficiency Adenosine DeAminase aciviy
infections
Frequent severe bacterial
Frequent severe viral infections
Frequent severe fungal infections
 Purine NucleosidePhosphorylase(PNP)
Deficiency

OH
o OH

12)hypoxanthino (4)a-0-ribose 1-phosphate

NH
ON oH human
PNP
OH
CHO
a OH
OH OH (2]hypoxanthino 3)o-riboso
inosine
1
ribooxacarbenium-ion
transitionstato

OH

OH OH
[5]N3-inosme isomer
 (SCID)
Adenosine Deaminase Deficiency
&Accumulation of dATP =inhibition of ribonucleotide

B andT
reductase
cells unable to divide
ADENOSINE DEAMINASE DEFICIENCY

INPURINE DEGRADATION, ADENosINE INosINE

ENZYME ISADA
ADA DEFICIENCY RESULTS IN SCID
"SEVERE COMBINED IMMUNODEFICIENCY"
SELECTIVELY KILLS LYMPHOCYTES
BOTH B-AND T-CELLS
MEDIATE MUCH OF IMMUNE RESPONSE
 In 1990 Ashanti de Silva became the
first patient to receive gene therapy for

ADA deficiency. Shown here at age13,

a healthy, active life
she continues to lead
1162023 Photo: of Van de Silua
 Pyrimidine Salvage
Uridine
phosphorylase
Uracil Ribose 1-phosphate Unridine+ Pi

Uridine UMP
Uridine-cytidine
or ATP Kinase
or ADP
Cytidine CMP

Thymidine
Thymine + Deoxyribose 1-phosphate Thymidine
Pi
phosphorylase

Thymidine+ ATP Thymidine

Kinase
dTMP ADP
Pyrimidine s
1162023
 Pyrimidine Degradation

to intermediates of
Pyrimidines are generally degraded
carbon metabolism (forexample, succinyl-CoA)
and
ammonia (NH).
Urea
NH4 ispackaged as urea through H,N-C-NH
the urea cycle and excreted by humans

Defects in of pyrimidine degradation have been

enzymes
documented, resulting in increased levels of pyrimidines
and neurological disorders.
 Class ACtVIty
6. Identify the Clinical Disorders/Diseases associated

with the metabolism of:

a) Purines b) Pyrimidines
Indicate theenzyme/s which is/are defectiveldeficient.

7. Identify the excretory products of

a) Purines b) Pyrimidines
8.What are anti-folate drugs? Why are they used in
cancer chemotheraphy?

162023
Adenosine Deaminase (ADA)Deficiency

ADA catalyses the ireversible hydrolytic deamination of

adenosine nucleosides to inosine nucleosides and
ammonia.

HO
Adenosine deaminase
NH

Adenylate deaminaso
0-P-0
ess
 Tetrahydrofolate (THE)

O Methylation of dUMP catalyzed by thymidylate synthase

Cofactor: N5,N10-methylene THF
Oxidized to dihydrofolate
Only known rxn where net oxidation state of THF changes
O THF Regeneration:
DHF+NADPH+ H*THF+NADP (enzyme:dihydrofolate
reductase)
THF+Serine Ns,N10-methy lene-THF + Glycine
(enzyme: serine hydroxymethyl transferase)

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 F H3CO NH2
HN
H3CO N
H NH2
Fluorouracil Trimethoprim

H2N

N
NH2
N CH2
10
çoO
N --NH-HCH2CH2Cop
CH3
Methotrexate
 Class Activity
1. What are the purine and pyrimidine bases?
2. In sequence, indicate the de novo biosynthesis of

a) Purine Nucleotides
b) Pyrimidine Nucleotides
3. Identify the precursors of the different atoms of :
a) Purine nucleus
b) Pyrimidine nucleus
4. Differentiate de novo biosynthesis from salvage
pathways.
5. Identify the regulatory control ofpurine nucleotide
biosynthesis.

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 dUMP dTMP

thymidylate
synthase

N5,N10-Methylene 7,8-Dihydrofoiate
Ha folate

serine
NADPH+ H*
Glycine hydroxymethyl-
transferase
dihydrofolate Methotrexate
reductase
PLP Aminopterin
Trimethoprim
H folate

Serine NADP
 Degradation of Pyrimidines

CMP and UMP degraded to bases similarly

to purines
Dephosphorylation
Deamination
Glycosidic bond cleavage

reduced in liver, forming B-alanine

Uracil
Converted tomalonyl-CoA fatty acid
synthesis for energy metabolism
 NH2
Nh2
H HN

Thymine 5-Methylcytosi ne
Cytosine Uracil

NADPH H Ring reduction

NADPH H
NADP NADP

HyTS

-1.Ring opens-
o
H CH3

Excreted r +2.C02 NH released

HaN-CH2-CH2-CoOH H2N-CH2-CH-COOH
Carnosine
Beta-alanine CH3 Excreted
Anserine Beto-omi noisobutyrate

16
2023
 NH3 NH3
CH2-C-c00
NH2 H2--coO
H
-cth
-CH2 -O
O= Glutamine Azaserine

Chemotherapeutic agents
often farget enzymes in NH3
the nucleotide
biosynthetic CH-G-COO
pathways...why?
-H2 H
CI
Acivicin
 NH2

H- NH4
HN HCH CH

Cytosine Uracil Thymine 5-Methylcytosi ne

NADPH H -Ring reduotion

NADPH H
NADP
NADP

HN -CH3

-1.Ring opens-
Excreted K -2C02 1Heleased
A2N-CH2-CH2-C00H H2N-CH2-CH-COOH
Cornosine
or Beta-alanine CH3 P Excreted
Anserine Beto-ami noisobutyrate

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 Anti-Folate Drugss

Cancer cells consume dTMP quickly for DNA replicatiunn

Interfere with thymidylate synthase reaction to
decrease dTMP production
(Fluorodeoxyuridylate- irreversible Inhibitor)
-also affects rapidly growing normal cells
(hair follicles, bone marrow, immune system
intestinal mucosa)
Dihydrofolate reductase step can
be stopped
competitively (DHF analogs)
Anti-Folates: Aminopterin, methotrexate,
trimethoprim
 NIWAUONO N 7
TETRAHYDROFOLATE
dUMP dTMP

NI-METH INL TUE

NADPH+H
GLYCINE
dilydro fola te redactase
seriae aydroxyme thy1
transferase
NADP
SERINE
 Jhanky
topeyou learned something

RAMON S. DEL FIERRO, Ph.D. (Tokyo)

Profe

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