Cloned (Comment) | Organism |
---|---|
expression of wild-type, selenomethinonine-labeled, and mutant R297A enzymes in Escherichia coli | Aeropyrum pernix |
Crystallization (Comment) | Organism |
---|---|
wild-type and selennomethionine-labeled enzyme, sitting drop method, 20 mg/ml protein in solution with 0.1 mM pyridoxal 5'-phosphate, 3 mM 2-mercaptoethanol, reservoir solution contains 0.1 M sodium cacodylate, pH 7.0, 0.2 M sodium acetate, 21% w/v poylethylene glycol 8000, and 3 mM 2-mercaptoethanol, mixture of equal volumes of protein and reservoir solution of 0.0035 ml, equilibration against 0.1 ml reservoir solution, 25°C, 2 weeks, crystallization of the selenomethinone enzyme at pH 6.0, multi-wave anomalous dispersion, X-ray diffraction structure determination and analysis at 2.0 A resolution | Aeropyrum pernix |
Protein Variants | Comment | Organism |
---|---|---|
R297A | site-directed mutagenesis, highly reduced activity with phospho-L-serine compared to the wild-type enzyme | Aeropyrum pernix |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
O-phospho-L-serine + hydrogen sulfide | Aeropyrum pernix | enzyme is involved in L-cysteine biosynthesis | L-cysteine + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aeropyrum pernix | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type, selenomethinonine-labeled, and mutant R297A enzymes from Escherichia coli | Aeropyrum pernix |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
O-phospho-L-serine + hydrogen sulfide = L-cysteine + phosphate | active site structure, modeling of substrate binding at the active site with Arg297 being crucial for activity | Aeropyrum pernix |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1 | - |
mutant R297A, substrate O-phospho-L-serine | Aeropyrum pernix |
53.1 | - |
mutant R297A, substrate O-acetyl-L-serine | Aeropyrum pernix |
72 | - |
wild-type enzyme, substrate O-acetyl-L-serine | Aeropyrum pernix |
245 | - |
wild-type enzyme, substrate O-phospho-L-serine | Aeropyrum pernix |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme also shows low L-cystathionine forming activity | Aeropyrum pernix | ? | - |
? | |
O-acetyl-L-serine + hydrogen sulfide | - |
Aeropyrum pernix | L-cysteine + acetate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | - |
Aeropyrum pernix | L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | enzyme is involved in L-cysteine biosynthesis | Aeropyrum pernix | L-cysteine + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | an enzyme monomer consists of 3 domains, structure overview | Aeropyrum pernix |
Synonyms | Comment | Organism |
---|---|---|
OPSS | - |
Aeropyrum pernix |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
85 | - |
assay at | Aeropyrum pernix |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | binding site in the cleft between middle and C-terminal domains through a covalent link to Lys127 | Aeropyrum pernix |