BRENDA - Enzyme Database
show all sequences of 2.5.1.65

Three-dimensional structure of a new enzyme, O-phosphoserine sulfhydrylase, involved in l-cysteine biosynthesis by a hyperthermophilic archaeon, Aeropyrum pernix K1, at 2.0 A resolution

Oda, Y.; Mino, K.; Ishikawa, K.; Ataka, M.; J. Mol. Biol. 351, 334-344 (2005)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of wild-type, selenomethinonine-labeled, and mutant R297A enzymes in Escherichia coli
Aeropyrum pernix
Crystallization (Commentary)
Crystallization
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
Engineering
Amino acid exchange
Commentary
Organism
R297A
site-directed mutagenesis, highly reduced activity with phospho-L-serine compared to the wild-type enzyme
Aeropyrum pernix
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
O-phospho-L-serine + hydrogen sulfide
Aeropyrum pernix
enzyme is involved in L-cysteine biosynthesis
L-cysteine + phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aeropyrum pernix
-
-
-
Purification (Commentary)
Commentary
Organism
recombinant wild-type, selenomethinonine-labeled, and mutant R297A enzymes from Escherichia coli
Aeropyrum pernix
Reaction
Reaction
Commentary
Organism
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 [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
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 and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme also shows low L-cystathionine forming activity
659728
Aeropyrum pernix
?
-
-
-
-
O-acetyl-L-serine + hydrogen sulfide
-
659728
Aeropyrum pernix
L-cysteine + acetate
-
-
-
?
O-phospho-L-serine + hydrogen sulfide
-
659728
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
O-phospho-L-serine + hydrogen sulfide
enzyme is involved in L-cysteine biosynthesis
659728
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
Subunits
Subunits
Commentary
Organism
More
an enzyme monomer consists of 3 domains, structure overview
Aeropyrum pernix
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
85
-
assay at
Aeropyrum pernix
Cofactor
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
binding site in the cleft between middle and C-terminal domains through a covalent link to Lys127
Aeropyrum pernix
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of wild-type, selenomethinonine-labeled, and mutant R297A enzymes in Escherichia coli
Aeropyrum pernix
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
binding site in the cleft between middle and C-terminal domains through a covalent link to Lys127
Aeropyrum pernix
Crystallization (Commentary) (protein specific)
Crystallization
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
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
R297A
site-directed mutagenesis, highly reduced activity with phospho-L-serine compared to the wild-type enzyme
Aeropyrum pernix
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
O-phospho-L-serine + hydrogen sulfide
Aeropyrum pernix
enzyme is involved in L-cysteine biosynthesis
L-cysteine + phosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type, selenomethinonine-labeled, and mutant R297A enzymes from Escherichia coli
Aeropyrum pernix
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
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 and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme also shows low L-cystathionine forming activity
659728
Aeropyrum pernix
?
-
-
-
-
O-acetyl-L-serine + hydrogen sulfide
-
659728
Aeropyrum pernix
L-cysteine + acetate
-
-
-
?
O-phospho-L-serine + hydrogen sulfide
-
659728
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
O-phospho-L-serine + hydrogen sulfide
enzyme is involved in L-cysteine biosynthesis
659728
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
an enzyme monomer consists of 3 domains, structure overview
Aeropyrum pernix
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
85
-
assay at
Aeropyrum pernix
Other publictions for EC 2.5.1.65
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
738193
Takeda
Role of F225 in O-phosphoserin ...
Aeropyrum pernix, Aeropyrum pernix DSM 11879
Extremophiles
20
733-745
2016
-
-
-
1
1
-
-
4
-
-
-
2
-
5
-
-
-
1
-
-
-
-
10
-
1
-
-
7
1
-
-
1
-
-
-
-
-
-
1
1
1
-
-
-
-
4
-
-
-
2
-
-
-
-
-
-
-
-
10
-
1
-
-
7
1
-
-
-
-
1
1
-
7
7
737868
Nakamura
Thermostability and reactivity ...
Aeropyrum pernix, Aeropyrum pernix DSM 11879
Biosci. Biotechnol. Biochem.
79
1280-1286
2015
-
-
1
-
-
-
-
-
-
-
-
2
2
6
-
-
1
1
-
-
-
-
6
-
1
-
1
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
1
-
-
-
-
6
-
1
-
1
-
1
-
-
-
-
1
1
-
-
-
738534
Steiner
CysK2 from Mycobacterium tuber ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Bacteriol.
196
3410-3420
2014
-
-
1
-
2
-
-
8
-
-
-
2
-
3
-
-
1
1
-
-
-
-
6
1
1
-
-
4
2
-
-
1
-
-
-
-
-
1
1
-
2
-
-
-
-
8
-
-
-
2
-
-
-
1
-
-
-
-
6
1
1
-
-
4
2
-
-
-
-
2
2
-
5
5
722990
Nakamura
Structural analysis of the sub ...
Aeropyrum pernix
J. Mol. Biol.
422
33-44
2012
-
-
1
1
11
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
1
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
685195
OLeary
O-Phospho-l-serine and the thi ...
Mycobacterium tuberculosis
Biochemistry
47
11606-11615
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
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-
2
-
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-
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-
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-
-
-
3
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
687773
Agren
Cysteine synthase (CYSM) of My ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
283
31567-31574
2008
-
-
1
1
1
-
-
-
-
-
-
4
-
6
-
-
1
-
-
-
-
-
8
-
-
-
-
1
-
-
-
1
-
-
-
-
-
2
2
2
2
-
-
-
-
-
-
-
-
4
-
-
-
1
-
-
-
-
8
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
702213
Jurgenson
Crystal structure of a sulfur ...
Mycobacterium tuberculosis
Biochemistry
47
10354-10364
2008
-
-
1
1
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
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-
1
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-
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-
1
1
1
1
-
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-
-
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-
-
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-
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-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
659728
Oda
Three-dimensional structure of ...
Aeropyrum pernix
J. Mol. Biol.
351
334-344
2005
-
-
1
1
1
-
-
-
-
-
-
1
-
7
-
-
1
1
-
-
4
-
4
1
1
-
-
-
-
-
-
1
-
-
-
-
-
1
1
1
1
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
4
-
4
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
637385
Mino
Characterization of a novel th ...
Aeropyrum pernix
J. Bacteriol.
185
2277-2284
2003
-
-
1
-
-
-
-
3
-
-
2
1
-
1
-
-
1
-
-
-
1
1
4
1
-
-
1
2
1
-
4
1
-
-
-
-
-
1
1
-
-
-
-
-
-
3
-
-
2
1
-
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1
-
-
1
1
4
1
-
-
1
2
1
-
4
-
-
-
-
-
-
-
654085
Mino
Crystallization and preliminar ...
Aeropyrum pernix
Acta Crystallogr. Sect. D
59
338-340
2003
-
-
1
1
-
-
-
-
-
-
-
3
-
1
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1
1
-
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4
1
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-
1
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1
1
1
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3
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1
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4
1
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-
-
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655576
Mino
A novel O-phospho-L-serine sul ...
Aeropyrum pernix
FEBS Lett.
551
133-138
2003
1
-
1
-
-
-
23
10
-
-
1
3
-
1
-
-
1
1
-
-
1
-
13
1
3
-
1
3
1
-
-
1
-
-
-
1
-
1
1
-
-
-
-
23
-
10
-
-
1
3
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-
-
1
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1
-
13
1
3
-
1
3
1
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