Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2(DE3)pLysS | Homo sapiens |
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 | 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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
214500 | - |
recombinant His-tagged R303C mutant enzyme | Homo sapiens |
225000 | - |
recombinant His-tagged wild-type enzyme | Homo sapiens |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P30566 | - |
- |
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 | Comment | Organism | Textmining |
---|---|---|---|
fibroblast | - |
Homo sapiens | - |
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 | Comment | Organism |
---|---|---|
homotetramer | recombinant His-tagged wild-type and R303C mutant enzymes, static light scattering | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
ADSL | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
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 |