Supplement for Ch18

Metabolic acidosis

Ammonia intoxication

Urea cycle defect

Vit B12 and folate

H4-biopterin

1
 Metabolic acidosis (p. 681)

Kidney extracts little Gln from bloodstream normally

Acidosis increases glutamine processing in kidney
 NH4+ + metabolic acids  salts (excreted in urine)
 -ketoglutarate  bicarbonate (HCO3-, buffer)
In kidney
Gln

TCA
cycle (buffer)
-ketoglutarate HCO3-
kidneys +
mitochondria NH4+ + acids Salts
(excreted)
Glutamate
dehydrogenase

Glu Fig 18-8 modified 2

NH4+ intoxication (p.681-682)

Build-up of urea cycle intermediates (Life threatening !!)
 Clinical symptoms:
 Increase [NH4+] in blood and urine
 Nausea and illness after ingestion of proteins
 Gradual mental retardation if not treated
 Coma, cerebral edema, increase cranial pressure

Possible mechanisms
 Depletion of ATP in brain cells
 Changes of cellular osmotic balance in brain
 Depletion of neurotransmitter

Remove excess NH4+
 Glutamate dehydrogenase: NH4+ + -KG  Glu
 Glutamine synthetase: NH4+ + Glu  Gln

[NH4+]  [Gln]  H2O uptake  cell swelling

[Glu]  [GABA]

[-KG]  ATP generated from citric acid cycle 3

 Defect in NH4+ removal

Treatments
 Strict diet control and supplements of essential a.a.
 With the administration of :
 Aromatic acids (Fig 18-14)
 Lower NH4+ level in blood
 (Aromatic acids) benzoate + Gly +  hippurate (left)
 Phenylbutyrate + Glutamine +  phenylacetylglutamine (right)
 Carbamoyl glutamate (N-acetylglutamate analog, Fig 18-13)
 Deficiency of N-acetylglutamate synthase
 Arginine (p.687)
 Deficiency of ornithine transcarbamoylase
 Deficiency of argininosuccinate synthetase
 Deficiency of argininosuccinase
 -keto acids mixtures (Concepts in Biochemistry, 2nd ed., p.541)
 Pick up excess NH4+ and converted to essential a.a.
 -keto--methylvalerate, -ketoisovalerate, -ketoisocaproate
 (Ile) (Val) (Leu)
4
Coming back to the essentials

From oxaloacetate and pyruvate
6 essential a.a. (Fig 22-15, from bacteria)
p. 865

5
From OAA and pyruvate

In bacteria (Fig 22-15 simplified)
Oxaloacetate Pyruvate p. 866-7

Aspartate Threonine -ketobutyrate

Aspartate - Homoserine -keto-

semialdehyde -keto-- isovalerate
methylvalerate

-keto-
Lysine Methionine Valine isocaproate

Isoleucine
Leucine
6
 Vit B12 and folate (p. 691)

Met synthesis in mammal
 N5-methyl H4 folate as C donor
 C is transferred to cobalamin derived from
Vit B12 (coenzyme B12)
 Vit B12 as the final C donor

Vit B12 deficiency
 Q: 18-12, 13 and 22-10
 H4 folate is trapped in N5-methyl form
 N5-methyl H4 folate is formed irreversibly
(Fig 18-17, top)
 Available folate
 e.g. pernicious anemia
 Biosynthesis of Gly  porphyrin  Hb

Fig 18-18 left 7

 p.690

FIGURE 1817 Conversions of

1-C units on tetrahydrofolate.
 H4 biopterin (p. 697-8)

H4 biopterin is required for
 Phe hydroxylation
 Formation of L-dopa  Dopamine Norepinephrine epinephrine
 Formation of 5-OH-Trp  serotonin

Dihydrobiopterin reductase is required to regenerate H4 biopterin
 Defect in dihydrobiopterin (H2 biopterin) reductase
 PKU + L-dopa, norepinephrine, serotonin deficiency
 Supplement with H4 biopterin (cant cross BBB), as well as 5-OH-Trp and L-dopa

Fig 18-24

9
Supplement for Ch22
Defect in purine metabolism
Folic acid and dT
Targets of chemotherapy
Gout and allopurinol

10
Defect in purine metabolism

Adenosine deaminase (ADA) deficiency P. 892-3
 100-fold increase in the cellular [dATP]
 Inhibit ribonucleotide reductase (Fig 22-42)
 A general deficiency of other dNTPs
 T and B lymphocytes do not develop properly
 Severe immunodeficiency disease
 A sterile bubble environment AMP
 Targets for human gene therapy trials (Box 9-2)
P

Lesch-Nyhan syndrome
GMP Adenosine
 Deficiency of purine salvage enzyme HGPRT
 Hypoxanthine-guanine phosphoribosyltransferase ADA
 [PRPP] and de novo synthesis [purine] P Inosine
 [uric acid] Guanosine PRPP
Ribose
 Gout-like tissue damage HGPRT Hypoxanthine
 Damage to CNS
PRPP
Ribose
Guanine Xanthine

Modified Fig 22-45 11

Uric acid
Folic acid and dT (p. 891)

Folic acid deficiency
 Heart disease, brain dysfunction and cancer
 Part arise from a reduction of dT synthesis
 Abnormal incorporation of U into DNA
 U is recognized by DNA repair pathways Fig 22-44/49
 Strand breaks

1-C transfer

Fig 22-43

12
 Targets of chemotherapy

The only cellular pathway for dT synthesis
 Thymidylate synthase
 Fluorouracil  FdUMP Fig 22-49
(mechanism-based inhibitor)
 Dihydrofolate reductase
 Methotrexate
 Folate analog, competitive I
 Aminopterin
 Competitive I
 Trimethoprim (antibiotics)
 Higher affinity for
bacterial enzyme

P. 894-6
13
 Problem 16 (Ch 22), p.898

Allopurinol is used to treat chronic gout
 Patients treated with allopurinol sometimes develop xanthine
stones in the kidneys
 The incidence of kidney damage is much lower than in
untreated gout.

Solubility in urine AMP
 Uric acid: 0.15 g/L GMP P
 Xanthine: 0.05 g/L
 Hypoxanthine: 1.4 g/L P
Guanosine
Ribose
Hypoxanthine
Ribose
Xanthine
Guanine Xanthine
oxidase

Uric acid
14
, , Why ?

Why we get sick ? Purine catabolism
Fig 22-45 right
 The new science of
Darwinian medicine Uric acid
Primates, birds,
reptiles, insects


 Uric acid Allantoin Most mammals

 Antioxidant
 Aging Allantoate Bony fishes
 Gout
 Longevity Amphibians,
Urea cartilaginous
fishes

Marine
NH4+ invertebrates
15