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Literature summary for 2.5.1.65 extracted from

  • Takeda, E.; Kunimoto, K.; Kawai, Y.; Kataoka, M.; Ishikawa, K.; Nakamura, T.
    Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1 (2016), Extremophiles, 20, 733-745.
    View publication on PubMed

Crystallization (Commentary)

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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Aeropyrum pernix Q9YBL2
-
-
Aeropyrum pernix DSM 11879 Q9YBL2
-
-

Reaction

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 and Products (Substrate)

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

Synonyms Comment Organism
ApOPSS
-
Aeropyrum pernix

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
assay at Aeropyrum pernix

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Aeropyrum pernix

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate linked to lysine K127 as an internal Schiff base at the active site Aeropyrum pernix

General Information

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 [mM/s]

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