NucleotideMetabolism Nucleotide Metabolism

Dr.AtifH.Khirelsied FacultyofMedicine InternationalUniversityofAfrica,Khartoum,Sudan I i lU i i f Af i Kh S d

OverviewofNucleotideMetabolism Overview of Nucleotide Metabolism

Nucleotidefunctions

Buildingblocksofnucleicacids. Energyforcellularprocesses(ATP). Biosyntheticintermediates(UTPGlc,CTPglycerol). Donorsofphosphoryl groups(phosphorylation Donors of phosphoryl groups (phosphorylation Rxns). Componentsofcofactors(NAD,FAD,FMNN,SAM). Cellularsignaling(2 messengers). Cellular signaling (2nd messengers) Allosteric regulatorsofenzymeactivity(PFK1).

OverviewofNucleotideMetabolism Overview of Nucleotide Metabolism

Biosyntheticpathways 1. denovosynthesis ofpurines andpyrimidinesis theprimarysource. the primary source. 2. salvagepathways(recycling) 2 salvage pathways (rec cling)
Phosphoribosylation ofbaseswithPRPP. Phosphorylation ofnucleosides

denovosynthesisofnucleotides de novo synthesis of nucleotides

Itisthesynthesisofnucleotidesfromsimpleprimary precursors. It is complex process requiring several precursors and Itiscomplexprocessrequiringseveralprecursorsand energyexpenditure.

PrecursorsforNucleotideBiosynthesis Precursors for Nucleotide Biosynthesis

5phosphoribosyl1pyrophosphate (PRPP) 5 phosphoribosyl 1 pyrophosphate(PRPP) ribosephosphatepyrophosphokinase Ribose5phosphate+ATP5phosphoribosyl1 pyrophosphate + AMP +AMP

PRPP

PrecursorsforNucleotideBiosynthesis y
Tetrahydrofolate (H4 folate) derivatives Tetrahydrofolate(H folate)derivatives
N5,N10methyleneH4 folate, N5,N10methenylH4 folate N5formylH4 folate thymidylate biosynthesis purinebiosynthesis purinebiosynthesis

PrecursorsforNucleotideBiosynthesis

PrecursorsforNucleotideBiosynthesis Precursors for Nucleotide Biosynthesis

Amino acids Aminoacids Glycineforpurinebiosynthesis A Aspartate f forpyrimidinebiosynthesis i idi bi h i

Aminoacidnitrogen g aminogroupofaspartate (purines) amidegroupofglutamine(purines,pyrimidines)

BiosynthesisofthePurineRing Biosynthesis of the Purine Ring

Multistepsynthesisfrommanyprecursors u t step sy t es s o a y p ecu so s

5 6

2 3

7 4 1

(numbersindicateorderofadditiontopurine ringfromPRPP)

Denovosynthesisofpurines

Step1

PRPPiscentraltovariouspathways

PurineBiosynthesis Purine Biosynthesis

Step2 St 2

glutaminedonatesamidenitrogentoactivated5 l i d id i i d5 phosphoribose(PRPP) committed step for purine synthesis committedstepforpurinesynthesis

PurineBiosynthesis Purine Biosynthesis

Step3 St 3

GARsynthetase glycine carboxylactivatedwithATP Pi displaced;amidebondformed

PurineBiosynthesis Purine Biosynthesis

Step4 St 4

GARtransformylase N10formyltetrahydrofolatedonatesformyl grouptoglycine aminogroup

PurineBiosynthesis Purine Biosynthesis

Step5 St 5

FGARamidotransferase FGAR amidotransferase ATPactivatescarbonylgroup amidotransferase displaces Pi displacesPi

PurineBiosynthesis Purine Biosynthesis

Step6 St 6

FGAMcyclase (AIRsynthetase) FGAM cyclase (AIR synthetase) ATPactivatescarbonyl cyclization of imidazole ring ofimidazole

PurineBiosynthesis Purine Biosynthesis

2steps

Step7 St 7
inhighereukaryotes: i hi h k t

inmicroorganisms:

AIRcarboxylase f formationofonlyCCbond f l b d nocofactorsorATPrequired

PurineBiosynthesis

Step8 St 8

SAICARsynthetase SAICAR synthetase aspartate isaminodonor ATP activates carboxylate ATPactivatescarboxylate aspartate aminoreplacesPi

PurineBiosynthesis

Step9 St 9
( (ADENYLOSUCCINASE) )

SAICARlyase SAICAR lyase fumarate iseliminated steps8and9analogoustoureacycle steps 8 and 9 analogous to urea cycle AICARfromhistidine biosynthesis

PurineBiosynthesis Purine Biosynthesis

Step10 St 10
( (FORMYLTRANSFERASE) )

AICARtransformylase N10formylH4 folate donatesformyl groupto glutaminederivedamine

PurineBiosynthesis Purine Biosynthesis

Step11 St 11

(IMPCYCLOHYDROLASE)

IMPsynthase cyclization ofsecondpurinering ATPactivationnotrequired

SynthesisofAdenylate andGuanylate
Step13 Step12

Step14 Step15

OrganizationofPurineBiosyntheticEnzymes
Purine nucleotides synthesis is carried out by multienzyme nucleotidessynthesisiscarriedoutbymultienzyme complexes Ineukaryotes,therearemultifunctionalproteinsfor: Steps 1 3 and 5 Steps1,3and5 Steps6and8 Steps10and11 I b Inbacteria,separateenzymesassociateinlargecomplexes i i i l l
Multienzymes affordefficientsynthesis,channeling,and y y , g, regulation.

RegulationofPurine Biosynthesis
FeedbackInhibition F db k I hibi i Inhibitionof1st stepbyADP Inhibitionof2nd stepbyIMP, AMPandGMP Inhibitionatthebranchpoint

SalvagePathwaysforNucleotides
SalvagepathwayaddscompletedpurinebasetoPRPP. Adenosine phosphoribosyltransferase de os e p osp o bosy t a sfe ase Adenine+PRPPAMP+ppi Hypoxanthineguaninephosphoribosyltransferase (HGPRT) Hypoxanthine+PRPPIMP+PPi Guanine+PRPPGMP+PPi LeschNyhan syndrome =LackofHGPRT activity. y y y

SalvagePathwaysforNucleotides

SalvagePathwaysforNucleotides

PyrimidineNucleotideSynthesis
Dr.AtifH.Khirelsied DepartmentofBiochemistry FacultyofMedicineandHealthSciences F lt f M di i d H lth S i InternationalUniversityofAfrica,Khartoum,Sudan

PyrimidineBiosynthesis y y

Precursorsofpyrimidinenucleotides

PyrimidineBiosynthesis Pyrimidine Biosynthesis

Step1 Step 1

Carbamoyl Ph h t S th t C b l PhosphateSynthetaseII II

Carbamoyl PhosphateSynthetaseII Cytosolic CPSIIenzyme(pyrimidines). Mit h d i l CPS I ( MitochondrialCPSI(urea) ) Bacteriahavesingleenzymeforbothfunctions

PyrimidineBiosynthesis Pyrimidine Biosynthesis

Step1: p p y ( ) 1. bicarbonatephosphatesynthesis(1st activation) 2. carbamate synthesis(NH3 fromglutaminehydrolysis) 3. carbamoyl phosphate synthesis (2nd activation) phosphatesynthesis(2

PyrimidineBiosynthesis Pyrimidine Biosynthesis

Step2

Step3,

Step4

Step5

PyrimidineBiosynthesis Pyrimidine Biosynthesis

Step6

Step7

Step8 St 8

Cytidine 5triphosphate(CTP)

ConversionofdUMP todTTP Conversion of dUMP to dTTP

Thymidylate synthase usesN5,N10 MethyleneH4 folate asbothone carbonsourceandreducingagent carbon source and reducing agent Dihydrofolate reductasereducesH2 y folate H4 folate withNADPH Serinehydroxymethyl transferase reactionrestores N5,N10 Methylene H MethyleneH4 folate

RegulationofPyrimidineBiosynthesis Regulation of Pyrimidine Biosynthesis

Feedbackinhibitionof1st stepaspartate transcarbamoylase (ATCase)byCTP ATPreverseseffectofCTP.

NucleotideBiosynthesis
PurineBiosynthesis
Hypoxanthine(apurine)is assembledontheribose5 phosphate phosphate Inosinate (IMP) (IMP) Precursors: PRPP Glycine H4 folateformate (2) HCO3 Glutamine(amideN)(2) A Aspartate ( i N) (aminoN) IMP AMP IMP XMP GMP IMP XMP

PyrimidineBiosynthesis
Orotate (apyrimidine)ismade firstthenaddedtoribose5 phosphate phosphate Orotidylate Precursors: Carbamoyl phosphate HCO3 Glutamine(amideN) Aspartate PRPP Orotidylate UMPUDP UTP CTP

CatabolismofNucleotides
Dr.Atif H.Khirelsied DepartmentofBiochemistry FacultyofMedicine,InternationalUniversityofAfrica y , y

Catabolismof PurineNucleotides
Adenosinedeaminase deficiency: Severe immunodeficiency disease; Severeimmunodeficiencydisease; lossofT andBcells 100xdATP (inhibitorof ribonucleotide reductase) ib l tid d t ) dNTPs,DNAsynthesis

CatabolismofPurineNucleotides
Catabolite of purines ofpurines.
Allopurinol

Gout hyperuricemia.

Uricacid

Allopurinol inhibits xanthine oxidase p

Purine eandPyrimidineC Catabo olism

Disordersofnucleotidesmetabolism Di d f l tid t b li
Gout
VariousgeneticdefectsinPRPPsynthetase present clinicallyasgout. clinically as gout
Eachdefect(e.g., Vmax, affinityforribose5 phosphate,orresistancetofeedbackinhibition) productionandexcretionofuricacid. production and excretion of uric acid

Hyperuricemia
Naurate crystallizes in soft tissues and joints leading to crystallizesinsofttissuesandjointsleadingto inflammatoryreaction,goutyarthritis.

Disordersofnucleotidesmetabolism Disorders of nucleotides metabolism

LeschNyhan Syndrome
Hyperuricemia LackofHGPRTactivity. Lack of HGPRT activity Episodesofuricacidlithiasis Bizarresyndromeofselfmutilation. Mentalretardation. Mental retardation Earlydeath

Disordersofnucleotidesmetabolism Disorders of nucleotides metabolism

VonGierkes Disease
Glucose6phosphatasedeficiency SecondarytoincreasedR5PandPRPP. Secondary to increased R5P and PRPP Associatedlacticacidosiselevatestherenalthreshold forurate.

Disordersofnucleotidesmetabolism Disorders of nucleotides metabolism

Adenosinedeaminase deficiency Adenosine deaminase deficiency
Associatedwithanimmunodeficiencydisease Dysfunctionallymphocytes(T andBcells).

Purine nucleosidephosphorylase deficiency AssociatedwithaseveredeficiencyofTcellsbutapparently normalBcellfunction.

ImmunedysfunctionsresultfromaccumulationofdGTP anddATP,which inhibitribonucleotide inhibit ribonucleotide reductase and thereby deplete cells of DNA andtherebydepletecellsofDNA precursors.

Catabolismofpyrimidinesnucleotides Catabolism of pyrimidines nucleotides

Productsofpyrimidinecatabolismarehighlywater soluble:(CO NH alanine and aminoisobutyrate) soluble: (CO2,NH3,alanine,andaminoisobutyrate).

Excretionofaminoisobutyrate increasesinleukemia E cretion of aminoisob t rate increases in le kemia andseverexrayradiationexposureduetoincreased destructionofDNA.

Catabolismof pyrimidinesnucleotides i idi l id

Catabolismofpyrimidinesnucleotides

End