1. Principles of Biochemistry
Horton • Moran • Scrimgeour • Perry • Rawn
Principles of Biochemistry
Fourth Edition
Chapter 18
Nucleotide Metabolism
Copyright © 2006 Pearson Prentice Hall, Inc.

2. Chapter 18 - Nucleotide Metabolism
Nucleotides are building blocks of DNA and RNA
Nucleotides are cosubstrates (ATP) and regulatory compounds (cyclic AMP)
Every nucleotide contains either a purine base or a pyrimidine base
This chapter presents the biosynthesis and degradation of nucleotides

3. 18.1 Synthesis of Purine Nucleotides
Fig 18.1 Uric acid and the major purines

4. Fig 18.2 Sources of ring atoms in purines synthesized de novo
The purine ring structure is not synthesized as a free base but as a substituent of ribose 5-phosphate.

5. Fig 18.3 Synthesis of 5-ribosyl 1-pyrophosphate (PRPP)

6. Fig 18.4 Inosine 5’-monophosphate (IMP or inosinate)
IMP is the initial product of the 10-step purine nucleotide pathway
Hypoxanthine (6-oxopurine) is the base for IMP

7. Fig 18.5 Ten-step pathway for de novo synthesis of IMP

8. Fig 18.5 Ten-step pathway for the synthesis of IMP (10 slides)

18. Fig 18.6 Mechanism for CO2 addition
Carboxyaminoimidazole ribonucleotide is an intermediate in the conversion of AIR to CAIR.

19. 18.2 Other Purine Nucleotides Are Synthesized from IMP
IMP can be converted to either AMP or GMP
Two enzymatic reactions are required for each
AMP and GMP can then be phosphorylated to their di- and triphosphates by kinases

20. Fig 18.7
Pathway for conversion of IMP to AMP and to GMP

21. Fig 18.7
Pathway for conversion of IMP to AMP and to GMP (continued next slide)

22. Fig 18.7 (continued)

23. Fig 18.8
Feedback inhibition in purine nucleotide biosynthesis

24. 18.3 Synthesis of Pyrimidine Nucleotides
Sources of the ring atoms in pyrimidines
Figure The immediate precursor of C-2 and N-3 is carbamoyl phosphate

25. The Major Pyrimidines

26. A. The Pathway for Pyrimidine Synthesis
Uridine 5’-phosphate is the precursor of other pyrimydine nculeotides
A six-step pathway to UMP
In eukaryotes, Steps 1-3 are catalyzed by a multifunctional protein (dihydroorotate synthase)
Steps 5,6 are catalyzed by a bifunctional enzyme (UMP synthase)

27. Six-step pathway for the de novo synthesis of UMP in prokaryotes
Fig 18.10
Six-step pathway for the de novo synthesis of UMP in prokaryotes

34. B. Regulation of Pyrimidine Synthesis
Eukaryotic cells have 2 carbamoyl phosphate synthetases (CPS I and CPS II)
Mitochondrial CPS I uses NH3 as nitrogen source

35. CPS II
Cytosolic CPS II uses glutamine as the nitrogen donor to carbamoyl phosphate

36. Regulation of pyrimidine synthesis
CPS II is allosterically regulated: PRPP and IMP are activators Several pyrimidines are inhibitors
Aspartate transcarbamoylase (ATCase) Important regulatory point in prokaryotes Catalyzes the first committed pathway step Allosteric regulators:
CTP (-), CTP + UTP (-), ATP (+)

37. Fig 18.11 Complex structure of ATCase from E. coli

38. 18.4 CTP is Synthesized from UMP
UMP is converted to CTP in three steps
UMP first converted to UDP and UTP
(18-1)

39. Fig 18.12
Conversion of UTP to CTP by CTP synthetase

40. Fig 18.13
Regulation of pyrimidine nucleotide synthesis in E. coli

41. 18.5 Reduction of Ribonucleotides to Deoxyribonucleotides
2’-Deoxyribonucleoside triphosphates (substrates for DNA polymerase) synthesized by enzymatic reduction of ribonucleotides
Ribonucleotide diphosphate reductase (RDR):
ADP dADP GDP dGDP
CDP dCDP UDP dUDP

42. Fig 18.14 Reduction of ribonucleotide diphosphates

44. 18.6 Methylation of dUMP Produces dTMP
Deoxythymidylate (dTMP) (required for DNA synthesis) is formed from UMP in 4 steps
UMP UDP dUDP dUMP dTMP

45. Two routes for conversion of dUDP to dUMP
1. dUDP + ADP dUMP + ATP
2. dUDP + ATP dUTP dUMP + PPi
H2O
ADP

46. Fig 18.15
Synthesis of dTMP from dUMP (2 slides)

49. Salvage of thymidine to generate dTMP
(18.6)

50. 18.7 Salvage of Purines and Pyrimidines
During cellular metabolism or digestion, nucleic acids are degraded to heterocyclic bases
These bases can be salvaged by direct conversion to 5’-mononucleotides
PRPP is the donor of the 5-phosphoribosyl group
Recycling of intact bases saves energy (reduced nitrogen sources are scarce)

51. Fig 18.16
Breakdown of nucleic acids

52. Fig 18.17 Degradation and salvage of purines

53. Fig 18.18 Interconversions of purine nucleotides

54. Fig 18.19 Interconversions of pyrimidine nucleotides

55. 18.8 Purine Catabolism
Most free purines and pyrimidines are salvaged, however some are catabolized
Birds, reptiles and primates (including humans) convert purines to uric acid

56. Separation of base from (deoxy) ribose
Purine-nucleoside phosphorylase (PNP) separates the free purine base from the ribose (or deoxyribose)
PNP
(Deoxy)Nucleoside + Pi
Base + (Deoxy)-a-D-Ribose 1-phosphate

57. Fig 18.20 Breakdown of hypoxanthine and guanine to uric acid

58. Gout results from excess sodium urate
Gout is caused from overproduction or inadequate excretion of uric acid
Sodium urate is relatively insoluble and can crystallize in tissues
Gout can be caused by a deficiency of hypoxanthine-guanine phosphoribosyltransferase or defective regulation of purine biosynthesis

59. Allopurinol is a treatment for gout
Allopurinol is converted in cells to oxypurinol, an inhibitor of xanthine dehydrogenase
Allopurinol prevents high levels of uric acid
Hypoxanthine, xanthine are more soluble

60. Fig 18.21
Further catabolism of uric acid by other organisms

64. 18.9 The Purine Nucleotide Cycle in Muscle
During intense exercise, AMP deaminase converts AMP to IMP, releasing NH3
Decreasing AMP levels shifts the equilibrium of adenylate kinase to produce more ATP for muscle contraction
IMP can be recycled to AMP (Fig 18.7)
Purine nucleotide cycle shown on next slide

65. Purine Nucleotide Cycle
Net reaction for one turn of the cycle is:
Aspartate + GTP + H2O
Fumarate + GDP + Pi + NH3

66. 18.10 Pyrimidine Metabolism
Pyrimidine nucleotides are hydrolyzed to the nucleosides and Pi
Then thymine, uracil and (deoxy) ribose 1-phosphate are produced
Catabolism of the thymine and uracil bases ends with intermediates of central metabolism

67. Fig 18.23
Catabolism of uracil and thymine