BRENDA - Enzyme Database
show all sequences of 2.5.1.65

Structural analysis of the substrate recognition mechanism in O-phosphoserine sulfhydrylase from the hyperthermophilic archaeon Aeropyrum pernix K1

Nakamura, T.; Kawai, Y.; Kunimoto, K.; Iwasaki, Y.; Nishii, K.; Kataoka, M.; Ishikawa, K.; J. Mol. Biol. 422, 33-44 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Aeropyrum pernix
Crystallization (Commentary)
Crystallization
Organism
structures of the enzyme without acetate, the complex formed by the K127A mutant with the external Schiff base of pyridoxal 5'-phosphate with O-phosphoserine, and the complex formed by the K127A mutant with the external Schiff base of pyridoxal 5'-phosphate with O-acetylserine, to 2.1 A resolution. No significant difference is seen in the overall structure between the free and complexed forms of the enzyme. The side chains of T152, S153, and Q224 interact with the carboxylate of the substrate. The position of R297 is significantly unchanged in the complex of the K127A mutant with the external Schiff base, allowing enough space for an interaction with O-phosphoserine. The positively charged environment around the entrance of the active site including S153 and R297 is important for accepting negatively charged substrates
Aeropyrum pernix
Engineering
Amino acid exchange
Commentary
Organism
K127A
mutant is inactive for cysteine synthesis and does not form the alpha-aminoacrylate intermediate
Aeropyrum pernix
Q224A
0.04% of wild-type activity
Aeropyrum pernix
R297A
0.3% of wild-type activity
Aeropyrum pernix
R297E
11% of wild-type activity
Aeropyrum pernix
R297K
0.2% of wild-type activity
Aeropyrum pernix
S153A
0.4% of wild-type activity
Aeropyrum pernix
S153T
0.2% of wild-type activity
Aeropyrum pernix
T152A
0.2% of wild-type activity
Aeropyrum pernix
T152S
71% of wild-type activity
Aeropyrum pernix
T203A
36% of wild-type activity
Aeropyrum pernix
T203M
0.3% of wild-type activity
Aeropyrum pernix
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aeropyrum pernix
Q9YBL2
-
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
23
-
pH 7.5, 80░C
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 catalyzes the reaction of EC 2.5.1.47, cysteine synthase
722990
Aeropyrum pernix
?
-
-
-
-
O-phospho-L-serine + hydrogen sulfide
-
722990
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
Cofactor
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
crystallization data
Aeropyrum pernix
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Aeropyrum pernix
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
crystallization data
Aeropyrum pernix
Crystallization (Commentary) (protein specific)
Crystallization
Organism
structures of the enzyme without acetate, the complex formed by the K127A mutant with the external Schiff base of pyridoxal 5'-phosphate with O-phosphoserine, and the complex formed by the K127A mutant with the external Schiff base of pyridoxal 5'-phosphate with O-acetylserine, to 2.1 A resolution. No significant difference is seen in the overall structure between the free and complexed forms of the enzyme. The side chains of T152, S153, and Q224 interact with the carboxylate of the substrate. The position of R297 is significantly unchanged in the complex of the K127A mutant with the external Schiff base, allowing enough space for an interaction with O-phosphoserine. The positively charged environment around the entrance of the active site including S153 and R297 is important for accepting negatively charged substrates
Aeropyrum pernix
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
K127A
mutant is inactive for cysteine synthesis and does not form the alpha-aminoacrylate intermediate
Aeropyrum pernix
Q224A
0.04% of wild-type activity
Aeropyrum pernix
R297A
0.3% of wild-type activity
Aeropyrum pernix
R297E
11% of wild-type activity
Aeropyrum pernix
R297K
0.2% of wild-type activity
Aeropyrum pernix
S153A
0.4% of wild-type activity
Aeropyrum pernix
S153T
0.2% of wild-type activity
Aeropyrum pernix
T152A
0.2% of wild-type activity
Aeropyrum pernix
T152S
71% of wild-type activity
Aeropyrum pernix
T203A
36% of wild-type activity
Aeropyrum pernix
T203M
0.3% of wild-type activity
Aeropyrum pernix
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
23
-
pH 7.5, 80░C
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 catalyzes the reaction of EC 2.5.1.47, cysteine synthase
722990
Aeropyrum pernix
?
-
-
-
-
O-phospho-L-serine + hydrogen sulfide
-
722990
Aeropyrum pernix
L-cysteine + phosphate
-
-
-
?
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
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
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
-
-
-
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
-
-
1
1
-
-
-
-
4
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
3
-
-
-
1
-
-
-
-
4
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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
-
-
-
1
-
-
1
-
13
1
3
-
1
3
1
-
-
-
-
-
-
-
-
-