EC Number | Cloned (Comment) | Organism |
---|---|---|
2.7.9.3 | gene selD, DNA and amino acid sequence determination and analysis of enzymes from Escherichia coli strains DH5alpha-T1 AND BL21-Gold (DE3), that differ in position 14 and 197, and compared to the DNA sequence from Escherichia coli strain K-12, recombinant overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21-Gold (DE3) | Escherichia coli |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.7.9.3 | E197D | site-directed mutagenesis | Escherichia coli |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
2.7.9.3 | Mg2+ | required | Escherichia coli |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.7.9.3 | 37500 | - |
- |
Escherichia coli |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.9.3 | ATP + selenide + H2O | Escherichia coli | - |
AMP + selenophosphate + phosphate | - |
? | |
2.7.9.3 | ATP + selenide + H2O | Escherichia coli DH5alpha-T1 and BL21-Gold(DE3) | - |
AMP + selenophosphate + phosphate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.7.9.3 | Escherichia coli | - |
gene selD | - |
2.7.9.3 | Escherichia coli DH5alpha-T1 and BL21-Gold(DE3) | - |
gene selD | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.7.9.3 | recombinant wild-type and mutant enzymes from Escherichia coli strain BL21-Gold (DE3) by ammonium sulfate fractionation, dialysis, and anion exchange chromatography | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.9.3 | 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate + selenide + H2O | the compound is used as a synthetic analogue of the substrate ATP for the monitoring and quantitative analysis of the functional activity of SPS | Escherichia coli | 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-monophosphate + selenophosphate + phosphate | - |
? | |
2.7.9.3 | 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate + selenide + H2O | the compound is used as a synthetic analogue of the substrate ATP for the monitoring and quantitative analysis of the functional activity of SPS | Escherichia coli DH5alpha-T1 and BL21-Gold(DE3) | 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-monophosphate + selenophosphate + phosphate | - |
? | |
2.7.9.3 | ATP + selenide + H2O | - |
Escherichia coli | AMP + selenophosphate + phosphate | - |
? | |
2.7.9.3 | ATP + selenide + H2O | - |
Escherichia coli DH5alpha-T1 and BL21-Gold(DE3) | AMP + selenophosphate + phosphate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.7.9.3 | dimer | 4 * 37500, SDS-PAGE, in solution, the recombinant SPS exists as a dimer with two active sites capable of ATP binding in each subunit | Escherichia coli |
2.7.9.3 | tetramer | 4 * 37500, SDS-PAGE, the enzyme SPS seems to exist in a form of tetramer in given reaction conditions, in accordance with the concentration stoichiometry of 4 mol of TNP-ATP to 1 mole of recombinant protein, molecular structure modelling | Escherichia coli |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.7.9.3 | SelD | - |
Escherichia coli |
2.7.9.3 | selenophosphate synthetase | - |
Escherichia coli |
2.7.9.3 | SPS | - |
Escherichia coli |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
2.7.9.3 | 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate | i.e. TNP-ATP, binding analysis of recombinant wild-type and mutant E197D to TNP-ATP, a synthetic fluorescent nucleotide analogue of ATP. The compound is used as a synthetic analogue of the substrate ATP for the monitoring and quantitative analysis of the functional activity of SPS. A non-linear regression analysis of the saturation curve of TNP-ATP binding to D197 SPS fitting to a model with 2 distinct binding sites with KDs different in order. Kinetics, overview | Escherichia coli | |
2.7.9.3 | ATP | - |
Escherichia coli |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.7.9.3 | additional information | the non-linear regression analysis of the saturation curve of TNP-ATP binding to D197 SPS fits to a model with 2 distinct binding sites with KDs different in order. Enzyme SPS exists in a form of tetramer in given reaction conditions, in accordance with the concentration stoichiometry of 4 mol of 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate to 1 mole of recombinant protein | Escherichia coli |