Crystallization (Comment) | Organism |
---|---|
enzyme ApOPSS in complex with aminoacrylate intermediate formed from pyridoxal 5'-phosphate with O-phospho-L-serine or enzyme in complex with L-cysteine, hanging drop vapor diffusion method, mixing of 10 mg/ml protein in 50 mM potassium phosphate, pH 7.5, containing 0.2 mM pyridoxal 5'-phosphate, 2 mM EDTA, and 2 mM TCEP-HCl, with reservoir solutions containing 0.1 M HEPES, pH 7.5, 27% v/v 2-propanol, 10-12% v/v PEG 4000, and 12 mM TCEP-HCl, X-ray diffraction structure determinatoion and analysis at 2.14-2.15 A resolution, structure modeling | Aeropyrum pernix |
Protein Variants | Comment | Organism |
---|---|---|
F225A | site-directed mutagenesis, the Km value toward O-phospho-L-serine is not significantly different between the wild-type ApOPSS and the F225A mutant, the kcat value of the wild-type ApOPSS is 4.2fold higher toward O-phospho-L-serine and 15fold higher toward O-acetyl-L-erine than that of the F225A mutant, respectively. The mutation from phenylalanine to alanine at position 225 affects the catalytic activity, not substrate binding | Aeropyrum pernix |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics, the Km value toward O-phospho-L-serine is not significantly different between the wild-type ApOPSS and the F225A mutant, the kcat value of the wild-type ApOPSS is 4.2fold higher toward O-phospho-L-serine and 15fold higher toward O-acetyl-L-erine than that of the F225A mutant, respectively | Aeropyrum pernix | |
0.39 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-acetyl-L-serine | Aeropyrum pernix | |
1.6 | - |
hydrogen sulfide | pH 7.5, 80°C, mutant F225A, with O-phospho-L-serine | Aeropyrum pernix | |
3.8 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-phospho-L-serine | Aeropyrum pernix |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
O-phospho-L-serine + hydrogen sulfide | Aeropyrum pernix | - |
L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | Aeropyrum pernix DSM 11879 | - |
L-cysteine + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aeropyrum pernix | Q9YBL2 | - |
- |
Aeropyrum pernix DSM 11879 | Q9YBL2 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
O-phospho-L-serine + hydrogen sulfide = L-cysteine + phosphate | ping-pong bi-bi mechanism, the active site of ApOPSS contains pyridoxal 5'-phosphate linked to lysine 127 as an internal Schiff base. Binding of the primary substrate O-phospho-L-serine displaces the lysine and forms an external Schiff base, initiating the first half-reaction that yields an alpha-aminoacrylate intermediate linked to pyridoxal 5'-phosphate. The second half-reaction involves the addition of a secondary substrate to the alpha-aminoacrylate intermediate and generates an external Schiff base with cysteine. The active-site lysine reacts with this external Schiff base, releasing cysteine and regenerating the internal Schiff base with K127. When other nucleophiles are used instead of sulfide, enzyme ApOPSS produces the corresponding non-natural amino acid | Aeropyrum pernix |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is also active with O-acetyl-L-serine, cf. EC 2.5.1.47. When other nucleophiles are used instead of sulfide, enzyme ApOPSS produces the corresponding non-natural amino acid, when thiosulfate is used as nucleophile, for example, S-sulfocysteine is produced. In absence of sulfide, the primary substrate reacts with to pyridoxal 5'-phosphate in enzyme OPSS to yield an alpha-aminoacrylate intermediate, which is formed through an external Schiff base with the elimination of phosphate or acetate, the intermediate is finally degraded to pyruvate and pyridoxal 5'-phosphate by a water molecule without a nucleophile | Aeropyrum pernix | ? | - |
? | |
additional information | the enzyme is also active with O-acetyl-L-serine, cf. EC 2.5.1.47. When other nucleophiles are used instead of sulfide, enzyme ApOPSS produces the corresponding non-natural amino acid, when thiosulfate is used as nucleophile, for example, S-sulfocysteine is produced. In absence of sulfide, the primary substrate reacts with to pyridoxal 5'-phosphate in enzyme OPSS to yield an alpha-aminoacrylate intermediate, which is formed through an external Schiff base with the elimination of phosphate or acetate, the intermediate is finally degraded to pyruvate and pyridoxal 5'-phosphate by a water molecule without a nucleophile | Aeropyrum pernix DSM 11879 | ? | - |
? | |
O-acetyl-L-serine + hydrogen sulfide | low activity | Aeropyrum pernix | L-cysteine + acetate | - |
? | |
O-acetyl-L-serine + hydrogen sulfide | low activity | Aeropyrum pernix DSM 11879 | L-cysteine + acetate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | - |
Aeropyrum pernix | L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | best substrate | Aeropyrum pernix | L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | - |
Aeropyrum pernix DSM 11879 | L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + hydrogen sulfide | best substrate | Aeropyrum pernix DSM 11879 | L-cysteine + phosphate | - |
? | |
O-phospho-L-serine + thiosulfate | - |
Aeropyrum pernix | S-sulfocysteine + phosphate | - |
? | |
O-phospho-L-serine + thiosulfate | - |
Aeropyrum pernix DSM 11879 | S-sulfocysteine + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ApOPSS | - |
Aeropyrum pernix |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
assay at | Aeropyrum pernix |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.6 | - |
O-acetyl-L-serine | pH 7.5, 80°C, mutant F225A | Aeropyrum pernix | |
33 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-acetyl-L-serine | Aeropyrum pernix | |
38 | - |
O-acetyl-L-serine | pH 7.5, 80°C, wild-type enzyme | Aeropyrum pernix | |
130 | - |
hydrogen sulfide | pH 7.5, 80°C, mutant F225A, with O-phospho-L-serine | Aeropyrum pernix | |
150 | - |
O-phospho-L-serine | pH 7.5, 80°C, mutant F225A | Aeropyrum pernix | |
630 | - |
O-phospho-L-serine | pH 7.5, 80°C, wild-type enzyme | Aeropyrum pernix | |
730 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-phospho-L-serine | Aeropyrum pernix |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Aeropyrum pernix |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | linked to lysine K127 as an internal Schiff base at the active site | Aeropyrum pernix |
General Information | Comment | Organism |
---|---|---|
additional information | three-dimensional structure analysis of enzyme ApOPSS in complex with aminoacrylate intermediate formed from pyridoxal 5'-phosphate with O-phospho-L-serine or the enzyme in complex with L-cysteine, and structure comparisons, molecular docking simulation using the structure of PDB ID 3VSA as the wild-type enzyme ApOPSS, overview | Aeropyrum pernix |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
O-acetyl-L-serine | pH 7.5, 80°C, mutant F225A | Aeropyrum pernix | |
2.3 | - |
O-phospho-L-serine | pH 7.5, 80°C, mutant F225A | Aeropyrum pernix | |
3.5 | - |
O-acetyl-L-serine | pH 7.5, 80°C, wild-type enzyme | Aeropyrum pernix | |
11 | - |
O-phospho-L-serine | pH 7.5, 80°C, wild-type enzyme | Aeropyrum pernix | |
81 | - |
hydrogen sulfide | pH 7.5, 80°C, mutant F225A, with O-phospho-L-serine | Aeropyrum pernix | |
85 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-acetyl-L-serine | Aeropyrum pernix | |
190 | - |
hydrogen sulfide | pH 7.5, 80°C, wild-type enzyme, with O-phospho-L-serine | Aeropyrum pernix |