BRENDA - Enzyme Database

Expression, purification and kinetic characterisation of human tissue transglutaminase

Roy, I.; Smith, O.; Clouthier, C.M.; Keillor, J.W.; Protein Expr. Purif. 87, 41-46 (2013)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.3.2.13
expressed in Escherichia coli BL21(DE3) cells
Homo sapiens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.2.13
acetonitrile
48% residual activity at 5% (v/v)
Homo sapiens
2.3.2.13
acrylonitrile
68 and 48% residual activity at 2.5 and 5.0 mM, respectively
Homo sapiens
2.3.2.13
CP30a
reversible inhibitor
Homo sapiens
2.3.2.13
CP4d
reversible inhibitor
Homo sapiens
2.3.2.13
DMF
90% residual activity at 2.5 and 5% (v/v)
Homo sapiens
2.3.2.13
NC-I052
irreversible inhibitor
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.2.13
0.011
-
N-acetyl-PNPQLPF
apparent value, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.014
-
Ac-PNPQLPF-OH
at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.014
-
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C
Homo sapiens
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.3.2.13
Ca2+
dependent on
Homo sapiens
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.2.13
77600
-
x * 77600, calculated from SDS-PAGE; x * 77600, untagged enzyme, estimated from SDS-PAGE
Homo sapiens
2.3.2.13
104000
-
x * 104000, GST and PreScission Protease-tagged enzyme, estimated from SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.3.2.13
N-acetyl-PNPQLPF + alkylamine
Homo sapiens
-
?
-
-
?
2.3.2.13
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine + alkylamine
Homo sapiens
-
?
-
-
?
2.3.2.13
[protein]-L-glutamine + alkylamine
Homo sapiens
-
[protein]-N5-alkyl-L-glutamine + NH3
-
-
?
Organic Solvent Stability
EC Number
Organic Solvent
Commentary
Organism
2.3.2.13
dimethylformamide
about 90% activity at 2.5 and 5.0 mM dimethylformamide
Homo sapiens
2.3.2.13
DMSO
100% activity in the presence of 2.5 and 5.0 mM DMSO
Homo sapiens
2.3.2.13
additional information
the enzyme is not affected by DMSO
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.2.13
Homo sapiens
P21980
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.2.13
glutathione Sepharose 4B resin chromatography; glutathione Sepharose column chromatography
Homo sapiens
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.2.13
0.01
-
recombinant enzyme from lysate supernatant, at pH 7.0 and 25°C; unpurified enzyme, with N-Cbz-Glu(gamma-4-nitrophenylester)Gly as substrate, at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.33
-
purified enzyme, with N-Cbz-Glu(gamma-4-nitrophenylester)Gly as substrate, at pH 7.0 and 25°C; purified recombinant enzyme, at pH 7.0 and 25°C
Homo sapiens
Storage Stability
EC Number
Storage Stability
Organism
2.3.2.13
-80C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, several months, no loss of activity
Homo sapiens
2.3.2.13
4°C, lyophilised purified enzyme, 1 week, 80% loss of activity
Homo sapiens
2.3.2.13
4°C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, 1 week, 10% loss of activity
Homo sapiens
2.3.2.13
4°C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, 2 weeks, 25% loss of activity
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.2.13
Ac-PNPQLPF-OH + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
N-acetyl-PNPQLPF + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
N-Cbz-Glu(gamma-4-nitrophenylester)Gly + alkylamine
-
720978
Homo sapiens
4-nitrophenol + ?
-
-
-
?
2.3.2.13
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
[protein]-L-glutamine + alkylamine
-
720978
Homo sapiens
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.3.2.13
?
x * 104000, GST and PreScission Protease-tagged enzyme, estimated from SDS-PAGE; x * 77600, calculated from SDS-PAGE; x * 77600, untagged enzyme, estimated from SDS-PAGE
Homo sapiens
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.3.2.13
0.001
-
CP4d
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.004
-
CP30a
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.019
-
NC-I052
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.2.13
expressed in Escherichia coli BL21(DE3) cells
Homo sapiens
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.2.13
acetonitrile
48% residual activity at 5% (v/v)
Homo sapiens
2.3.2.13
acrylonitrile
68 and 48% residual activity at 2.5 and 5.0 mM, respectively
Homo sapiens
2.3.2.13
CP30a
reversible inhibitor
Homo sapiens
2.3.2.13
CP4d
reversible inhibitor
Homo sapiens
2.3.2.13
DMF
90% residual activity at 2.5 and 5% (v/v)
Homo sapiens
2.3.2.13
NC-I052
irreversible inhibitor
Homo sapiens
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.3.2.13
0.001
-
CP4d
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.004
-
CP30a
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.019
-
NC-I052
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C; at pH 7.0 and 25°C
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.2.13
0.011
-
N-acetyl-PNPQLPF
apparent value, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.014
-
Ac-PNPQLPF-OH
at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.014
-
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine
0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0 and 25°C
Homo sapiens
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.3.2.13
Ca2+
dependent on
Homo sapiens
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.2.13
77600
-
x * 77600, calculated from SDS-PAGE; x * 77600, untagged enzyme, estimated from SDS-PAGE
Homo sapiens
2.3.2.13
104000
-
x * 104000, GST and PreScission Protease-tagged enzyme, estimated from SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.3.2.13
N-acetyl-PNPQLPF + alkylamine
Homo sapiens
-
?
-
-
?
2.3.2.13
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine + alkylamine
Homo sapiens
-
?
-
-
?
2.3.2.13
[protein]-L-glutamine + alkylamine
Homo sapiens
-
[protein]-N5-alkyl-L-glutamine + NH3
-
-
?
Organic Solvent Stability (protein specific)
EC Number
Organic Solvent
Commentary
Organism
2.3.2.13
dimethylformamide
about 90% activity at 2.5 and 5.0 mM dimethylformamide
Homo sapiens
2.3.2.13
DMSO
100% activity in the presence of 2.5 and 5.0 mM DMSO
Homo sapiens
2.3.2.13
additional information
the enzyme is not affected by DMSO
Homo sapiens
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.2.13
glutathione Sepharose 4B resin chromatography; glutathione Sepharose column chromatography
Homo sapiens
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.2.13
0.01
-
recombinant enzyme from lysate supernatant, at pH 7.0 and 25°C; unpurified enzyme, with N-Cbz-Glu(gamma-4-nitrophenylester)Gly as substrate, at pH 7.0 and 25°C
Homo sapiens
2.3.2.13
0.33
-
purified enzyme, with N-Cbz-Glu(gamma-4-nitrophenylester)Gly as substrate, at pH 7.0 and 25°C; purified recombinant enzyme, at pH 7.0 and 25°C
Homo sapiens
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.3.2.13
-80C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, several months, no loss of activity
Homo sapiens
2.3.2.13
4°C, lyophilised purified enzyme, 1 week, 80% loss of activity
Homo sapiens
2.3.2.13
4°C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, 1 week, 10% loss of activity
Homo sapiens
2.3.2.13
4°C, purified enzyme, 0.05 mM EDTA, 3.3 mM CaCl2, in 0.11 M MOPS at pH 7.0, 2 weeks, 25% loss of activity
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.2.13
Ac-PNPQLPF-OH + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
N-acetyl-PNPQLPF + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
N-Cbz-Glu(gamma-4-nitrophenylester)Gly + alkylamine
-
720978
Homo sapiens
4-nitrophenol + ?
-
-
-
?
2.3.2.13
N-Cbz-L-glutaminyl(gamma-4-nitrophenylester)glycine + alkylamine
-
720978
Homo sapiens
?
-
-
-
?
2.3.2.13
[protein]-L-glutamine + alkylamine
-
720978
Homo sapiens
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.3.2.13
?
x * 104000, GST and PreScission Protease-tagged enzyme, estimated from SDS-PAGE; x * 77600, calculated from SDS-PAGE; x * 77600, untagged enzyme, estimated from SDS-PAGE
Homo sapiens