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Literature summary for 4.3.2.2 extracted from

  • Ray, S.P.; Deaton, M.K.; Capodagli, G.C.; Calkins, L.A.; Sawle, L.; Ghosh, K.; Patterson, D.; Pegan, S.D.
    Structural and biochemical characterization of human adenylosuccinate lyase (ADSL) and the R303C ADSL deficiency-associated mutation (2012), Biochemistry, 51, 6701-6713.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2(DE3)pLysS Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified wild-type apo-enzyme and enzyme mutant R303C, high throughput screening by sitting drop method, method optimization, mixing of 0.004 ml of protein solution with 0.004 ml of reservoir solution and equilibration against 0.5 ml of reservoir solution, with a precipitant gradient for the native enzyme of 18-28% w/v PEG 6000, 0.1 M Tris, pH 8.0, and 12.5 mM MgCl2 hexahydrate, and for enzyme mutant R303C, a precipitant gradient of 18-28% w/v PEG 8000, 0.1 M Tris, pH 8.5, and 12.5 mM spermine tetrahydrochloride, X-ray diffraction structure determination and analysis at 2.7 A and 2.6 A resolution, respectively Homo sapiens

Protein Variants

Protein Variants Comment Organism
R303C naturally occuring type II mutation, observed as a homozygous mutation in two unrelated patients, the mutant shows reduced activity compared to the wild-type enzyme, KM values of R303C enzyme mutant increase 4fold for phosphoribosylsuccinyl-aminoimidazole carboxamide compared to that of wild-type enzyme, for succinyladenosine monophosphate the change is almost negligible, substrate binding of the enzyme is latered compared to the wild-type Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information the enzyme shows cooperativity and does not follow simple Michaelis-Menten kinetics, kinetic analysis, overview Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
214500
-
recombinant His-tagged R303C mutant enzyme Homo sapiens
225000
-
recombinant His-tagged wild-type enzyme Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1-(5-phosphoribosyl)-4-(N-succinocarboxamide)-5-aminoimidazole Homo sapiens
-
5'-phosphoribosyl-5-amino-4-imidazolecarboxamide + fumarate
-
?
succinyladenosine monophosphate Homo sapiens
-
AMP + fumarate
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens P30566
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta 2(DE3)pLysS by nickel affinity chromatography, dialysis, cleavage of the His-tag through thrombin, and gel filtration, to homogeneity Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
fibroblast
-
Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-(5-phosphoribosyl)-4-(N-succinocarboxamide)-5-aminoimidazole
-
Homo sapiens 5'-phosphoribosyl-5-amino-4-imidazolecarboxamide + fumarate
-
?
additional information interaction between the enzyme and phosphoribosylsuccinyl-aminoimidazole carboxamide, isothermal titration calorimetry, overview. Modeling of AMPand phosphoribosylaminoimidazole carboxamide, and fumarate binding in the active site of wild-type and R303C mutant enzymes Homo sapiens ?
-
?
succinyladenosine monophosphate
-
Homo sapiens AMP + fumarate
-
?

Subunits

Subunits Comment Organism
homotetramer recombinant His-tagged wild-type and R303C mutant enzymes, static light scattering Homo sapiens

Synonyms

Synonyms Comment Organism
ADSL
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Homo sapiens

General Information

General Information Comment Organism
malfunction enzyme mutation R303C is involved in ADSL deficiency, a rare autosomal recessive disorder, which causes a defect in purine metabolism resulting in neurological and physiological symptoms. The R303C mutation potentially has a disproportional decrease in activity toward its substrates Homo sapiens
metabolism the enzyme executes two non-sequential steps in the de novo synthesis of AMP: the conversion of phosphoribosylsuccinyl-aminoimidazole carboxamide to phosphoribosylaminoimidazole carboxamide, which occurs in the de novo synthesis of IMP, and the conversion of adenylosuccinate to AMP, which occurs in the de novo synthesis of AMP and also in the purine nucleotide cycle, using the same active site Homo sapiens