BRENDA - Enzyme Database show

A novel, arsenite-sensitive E2 of the ubiquitin pathway: purification and properties

Klemperer, N.S.; Berleth, E.S.; Pickart, C.M.; Biochemistry 11, 6035-6042 (1989)

Data extracted from this reference:

Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.2.24
arsenite
specific for vicinal sulfhydryl sites in proteins, irreversible or very slow reversible, instead of other inhibition with arsenite which is usually rapidly reversed upon addition of competitive dithiol compounds
Oryctolagus cuniculus
2.3.2.24
casein
71 microM inhibited by 50%, with 79 microM cytochrome c, competitive inhibition
Oryctolagus cuniculus
2.3.2.24
iodoacetamide
reacts slowly, relative insenstitive to iodoacetamide. This property distinguishes E2230K from other E2 proteins
Oryctolagus cuniculus
2.3.2.24
lysozyme
139 microM inhibited by 90%, with 79 microM cytochrome c, competitive inhibition
Oryctolagus cuniculus
2.3.2.24
additional information
relatively insensitive to iodoacetamide inhibition
Oryctolagus cuniculus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.2.24
0.0013
-
[histone H3]-L-lysine
pH 7.0, 37°C, Vmax, 0.06 microM
Oryctolagus cuniculus
2.3.2.24
0.125
-
[cytochrome c]-L-lysine
pH 7.0, 37°C, Vmax, 0.12 microM
Oryctolagus cuniculus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.2.24
230000
-
gel fitration
Oryctolagus cuniculus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.2.24
Oryctolagus cuniculus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.2.24
purification about 7fold with 20% recovery, gel filtration, ammonium sulfate fractionation, FPLC anion exchange on a Mono Q coloumn, E2230K is completely separated from other E2 proteins by the gel filtration step
Oryctolagus cuniculus
Reaction
EC Number
Reaction
Commentary
Organism
2.3.2.24
[E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [acceptor protein]-L-lysine = [E1 ubiquitin-activating enzyme]-L-cysteine + [acceptor protein]-N6-monoubiquitinyl-L-lysine
the enzyme forms a labile ubiquitin adduct in the presence of E1, ubiquitin, and MgATP and catalyzes the conjugation of ubiquitin to protein substrates, independent of E3
Oryctolagus cuniculus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
2.3.2.24
reticulocyte
-
Oryctolagus cuniculus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.2.24
0.81
-
pH 7.0, 37°C, purified enzyme, in the presence of 79 microM yeast cytochrome c and 0.1 microM E1 protein
Oryctolagus cuniculus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.2.24
additional information
beta-lactoglobulin and reduced carboxymethylated bovine serum albumin rcmBSA are not conjugates
724254
Oryctolagus cuniculus
?
-
-
-
-
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [alpha-casein]-L-lysine
42% relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [alpha-casein]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [alpha-lactalbumin]-L-lysine
12% with human and 15% with bovine alpha-lactalbumin, relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [alpha-lactalbumin]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [cytochrome c]-L-lysine
yeast cytochrome c, ubiquitin from human blood
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [cytochrome c]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [histone H3]-L-lysine
-
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [histone H3]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [lysozyme]-L-lysine
5% relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [lysozyme]-N6-ubiquitinyl-L-lysine
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.3.2.24
monomer
1 * 230000, SDS-PAGE
Oryctolagus cuniculus
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.2.24
37
-
assay at
Oryctolagus cuniculus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.3.2.24
0.003
-
[histone H3]-L-lysine
pH 7.0, 37°C, Vmax, 0.06 microM
Oryctolagus cuniculus
2.3.2.24
0.00617
-
[cytochrome c]-L-lysine
pH 7.0, 37°C, Vmax, 0.12 microM
Oryctolagus cuniculus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.2.24
7
-
assay at
Oryctolagus cuniculus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.2.24
arsenite
specific for vicinal sulfhydryl sites in proteins, irreversible or very slow reversible, instead of other inhibition with arsenite which is usually rapidly reversed upon addition of competitive dithiol compounds
Oryctolagus cuniculus
2.3.2.24
casein
71 microM inhibited by 50%, with 79 microM cytochrome c, competitive inhibition
Oryctolagus cuniculus
2.3.2.24
iodoacetamide
reacts slowly, relative insenstitive to iodoacetamide. This property distinguishes E2230K from other E2 proteins
Oryctolagus cuniculus
2.3.2.24
lysozyme
139 microM inhibited by 90%, with 79 microM cytochrome c, competitive inhibition
Oryctolagus cuniculus
2.3.2.24
additional information
relatively insensitive to iodoacetamide inhibition
Oryctolagus cuniculus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.2.24
0.0013
-
[histone H3]-L-lysine
pH 7.0, 37°C, Vmax, 0.06 microM
Oryctolagus cuniculus
2.3.2.24
0.125
-
[cytochrome c]-L-lysine
pH 7.0, 37°C, Vmax, 0.12 microM
Oryctolagus cuniculus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.2.24
230000
-
gel fitration
Oryctolagus cuniculus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.2.24
purification about 7fold with 20% recovery, gel filtration, ammonium sulfate fractionation, FPLC anion exchange on a Mono Q coloumn, E2230K is completely separated from other E2 proteins by the gel filtration step
Oryctolagus cuniculus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
2.3.2.24
reticulocyte
-
Oryctolagus cuniculus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.2.24
0.81
-
pH 7.0, 37°C, purified enzyme, in the presence of 79 microM yeast cytochrome c and 0.1 microM E1 protein
Oryctolagus cuniculus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.2.24
additional information
beta-lactoglobulin and reduced carboxymethylated bovine serum albumin rcmBSA are not conjugates
724254
Oryctolagus cuniculus
?
-
-
-
-
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [alpha-casein]-L-lysine
42% relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [alpha-casein]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [alpha-lactalbumin]-L-lysine
12% with human and 15% with bovine alpha-lactalbumin, relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [alpha-lactalbumin]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [cytochrome c]-L-lysine
yeast cytochrome c, ubiquitin from human blood
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [cytochrome c]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [histone H3]-L-lysine
-
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [histone H3]-N6-ubiquitinyl-L-lysine
-
-
-
?
2.3.2.24
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [lysozyme]-L-lysine
5% relative activity compared to cytochrome c as substrate
724254
Oryctolagus cuniculus
[ubiquitin-carrier protein E2]-L-cysteine + [lysozyme]-N6-ubiquitinyl-L-lysine
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.3.2.24
monomer
1 * 230000, SDS-PAGE
Oryctolagus cuniculus
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.2.24
37
-
assay at
Oryctolagus cuniculus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.3.2.24
0.003
-
[histone H3]-L-lysine
pH 7.0, 37°C, Vmax, 0.06 microM
Oryctolagus cuniculus
2.3.2.24
0.00617
-
[cytochrome c]-L-lysine
pH 7.0, 37°C, Vmax, 0.12 microM
Oryctolagus cuniculus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.2.24
7
-
assay at
Oryctolagus cuniculus
General Information
EC Number
General Information
Commentary
Organism
2.3.2.24
physiological function
the enzyme forms a labile ubiquitin adduct in the presence of E1, ubiquitin, and MgATP and catalyzes the conjugation of ubiquitin to protein substrates, independent of E3
Oryctolagus cuniculus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.3.2.24
physiological function
the enzyme forms a labile ubiquitin adduct in the presence of E1, ubiquitin, and MgATP and catalyzes the conjugation of ubiquitin to protein substrates, independent of E3
Oryctolagus cuniculus