BRENDA - Enzyme Database show
show all sequences of 2.9.1.1

Structure and catalytic mechanism of eukaryotic selenocysteine synthase

Ganichkin, O.M.; Xu, X.M.; Carlson, B.A.; Mix, H.; Hatfield, D.L.; Gladyshev, V.N.; Wahl, M.C.; J. Biol. Chem. 283, 5849-5865 (2008)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of SecS as C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta2(DE3)
Mus musculus
Crystallization (Commentary)
Crystallization
Organism
purified recombinant C-terminally His6-tagged SecS, apoenzyme or in complex with phosphate or iodide, sitting-drop vapor diffusion at 20°C, 14 mg/ml protein in 10 mM HEPES-NaOH, pH 7.5, 500 mM NaCl, 2 mM DTT, is mixed with an equal volume of reservoir solution containing 11% v/v ethylene glycol without other buffer components, 2 days, cryoprotection by crystal soaking in 100 mM HEPES-NaOH, pH 7.5, 250 mM NaCl, 1 mM dithiothreitol, and 35% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 1.65-2.38 A resolution
Mus musculus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
20000
-
about, gel filtration
Mus musculus
49000
-
4 * 49000, SDS-PAGE
Mus musculus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-seryl-tRNASec + selenophosphate
Mus musculus
-
L-selenocysteinyl-tRNASec + phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mus musculus
-
-
-
Purification (Commentary)
Commentary
Organism
recombinant C-terminally His6-tagged SecS from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography and gel filtration to over 95% purity
Mus musculus
Reaction
Reaction
Commentary
Organism
L-seryl-tRNASec + selenophosphate = L-selenocysteinyl-tRNASec + phosphate
substrate binding and reaction mechanism
Mus musculus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-seryl-tRNASec + selenophosphate
-
693212
Mus musculus
L-selenocysteinyl-tRNASec + phosphate
-
-
-
?
L-seryl-tRNASec + selenophosphate
the phosphate loop accommodates the gamma-phosphate moiety of O-phospho-L-seryltRNA [Ser]Sec and, after phosphate elimination, binds selenophosphate to initiate attack on the proposed aminoacrylyltRNA [Ser]Sec intermediate. Binding of phosphate triggers disorder-order transition in an active site loop
693212
Mus musculus
L-selenocysteinyl-tRNASec + phosphate
-
-
-
?
additional information
SecS does not act on free O-phospho-L-serine
693212
Mus musculus
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
More
two SecS monomers interact intimately and together build up two identical active sites around PLP in a Schiff-base linkage with Lys284. Two SecS dimers further associate to form a homotetramer. The N-terminus, which mediates tetramer formation, and a large insertion that remodels the active site set SecS aside from other members of the family
Mus musculus
tetramer
4 * 49000, SDS-PAGE
Mus musculus
Cofactor
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
SecS is a a PLP-dependent transferase. The PLP cofactor iIs tightly anchored by non-canonical contacts to both protomers of a close dimer
Mus musculus
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of SecS as C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta2(DE3)
Mus musculus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
SecS is a a PLP-dependent transferase. The PLP cofactor iIs tightly anchored by non-canonical contacts to both protomers of a close dimer
Mus musculus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant C-terminally His6-tagged SecS, apoenzyme or in complex with phosphate or iodide, sitting-drop vapor diffusion at 20°C, 14 mg/ml protein in 10 mM HEPES-NaOH, pH 7.5, 500 mM NaCl, 2 mM DTT, is mixed with an equal volume of reservoir solution containing 11% v/v ethylene glycol without other buffer components, 2 days, cryoprotection by crystal soaking in 100 mM HEPES-NaOH, pH 7.5, 250 mM NaCl, 1 mM dithiothreitol, and 35% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 1.65-2.38 A resolution
Mus musculus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
20000
-
about, gel filtration
Mus musculus
49000
-
4 * 49000, SDS-PAGE
Mus musculus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-seryl-tRNASec + selenophosphate
Mus musculus
-
L-selenocysteinyl-tRNASec + phosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant C-terminally His6-tagged SecS from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography and gel filtration to over 95% purity
Mus musculus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-seryl-tRNASec + selenophosphate
-
693212
Mus musculus
L-selenocysteinyl-tRNASec + phosphate
-
-
-
?
L-seryl-tRNASec + selenophosphate
the phosphate loop accommodates the gamma-phosphate moiety of O-phospho-L-seryltRNA [Ser]Sec and, after phosphate elimination, binds selenophosphate to initiate attack on the proposed aminoacrylyltRNA [Ser]Sec intermediate. Binding of phosphate triggers disorder-order transition in an active site loop
693212
Mus musculus
L-selenocysteinyl-tRNASec + phosphate
-
-
-
?
additional information
SecS does not act on free O-phospho-L-serine
693212
Mus musculus
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
More
two SecS monomers interact intimately and together build up two identical active sites around PLP in a Schiff-base linkage with Lys284. Two SecS dimers further associate to form a homotetramer. The N-terminus, which mediates tetramer formation, and a large insertion that remodels the active site set SecS aside from other members of the family
Mus musculus
tetramer
4 * 49000, SDS-PAGE
Mus musculus
Other publictions for EC 2.9.1.1
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)
738683
Manhas
Leishmania donovani encodes a ...
Leishmania donovani
J. Biol. Chem.
291
1203-1220
2016
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725024
Manzine
Assembly stoichiometry of bact ...
Escherichia coli
FEBS Lett.
587
906-911
2013
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1
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3
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739586
Manzine
An efficient protocol for the ...
Escherichia coli
Protein Expr. Purif.
88
80-84
2013
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739706
Itoh
Decameric SelA-tRNA(Sec) ring ...
Aquifex aeolicus
Science
340
75-78
2013
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1
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693212
Ganichkin
Structure and catalytic mechan ...
Mus musculus
J. Biol. Chem.
283
5849-5865
2008
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1
1
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2
1
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5
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1
1
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1
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3
2
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691146
Fischer
Towards understanding selenocy ...
Moorella thermoacetica
Biol. Chem.
388
1061-1067
2007
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1
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1
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7
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691179
Xu
New developments in selenium b ...
Homo sapiens, Mus musculus
Biol. Trace Elem. Res.
119
234-241
2007
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2
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660828
Gursinsky
A selDABC cluster for selenocy ...
Peptoclostridium acidaminophilum
Arch. Microbiol.
174
200-212
2000
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1
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661290
Mizutani
Selenophosphate as a substrate ...
Bos taurus
Biofactors
9
27-36
1999
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1
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5
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22917
Tormay
Bacterial selenocysteine synth ...
Desulfomicrobium baculatum, Escherichia coli, Haemophilus influenzae, Moorella thermoacetica
Eur. J. Biochem.
254
655-661
1998
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2
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2
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8
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2
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22916
Tanabe
-
Selenophosphate as a substrate ...
Bos taurus
Nucleic acids Symp. Ser.
35
287-288
1996
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22915
Engelhardt
Structure of selenocysteine sy ...
Escherichia coli
Mol. Microbiol.
6
3461-3467
1992
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22912
Forchhammer
Selenocysteine synthase from E ...
Escherichia coli
J. Biol. Chem.
266
6324-6328
1991
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22913
Forchhammer
Selenocysteine synthase from E ...
Escherichia coli
J. Biol. Chem.
266
6318-6323
1991
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22914
Forchhammer
The function of selenocysteine ...
Escherichia coli
Biochimie
73
1481-1486
1991
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