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Literature summary for 2.3.2.24 extracted from

  • Klemperer, N.S.; Berleth, E.S.; Pickart, C.M.
    A novel, arsenite-sensitive E2 of the ubiquitin pathway: purification and properties (1989), Biochemistry, 11, 6035-6042.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
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
casein 71 microM inhibited by 50%, with 79 microM cytochrome c, competitive inhibition Oryctolagus cuniculus
iodoacetamide reacts slowly, relative insenstitive to iodoacetamide. This property distinguishes E2230K from other E2 proteins Oryctolagus cuniculus
lysozyme 139 microM inhibited by 90%, with 79 microM cytochrome c, competitive inhibition Oryctolagus cuniculus
additional information relatively insensitive to iodoacetamide inhibition Oryctolagus cuniculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0013
-
[histone H3]-L-lysine pH 7.0, 37°C, Vmax, 0.06 microM Oryctolagus cuniculus
0.125
-
[cytochrome c]-L-lysine pH 7.0, 37°C, Vmax, 0.12 microM Oryctolagus cuniculus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
230000
-
gel fitration Oryctolagus cuniculus

Organism

Organism UniProt Comment Textmining
Oryctolagus cuniculus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Reaction Comment Organism Reaction ID
[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

Source Tissue Comment Organism Textmining
reticulocyte
-
Oryctolagus cuniculus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information beta-lactoglobulin and reduced carboxymethylated bovine serum albumin rcmBSA are not conjugates Oryctolagus cuniculus ?
-
?
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [alpha-casein]-L-lysine 42% relative activity compared to cytochrome c as substrate Oryctolagus cuniculus [ubiquitin-carrier protein E2]-L-cysteine + [alpha-casein]-N6-ubiquitinyl-L-lysine
-
?
[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 Oryctolagus cuniculus [ubiquitin-carrier protein E2]-L-cysteine + [alpha-lactalbumin]-N6-ubiquitinyl-L-lysine
-
?
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [cytochrome c]-L-lysine yeast cytochrome c, ubiquitin from human blood Oryctolagus cuniculus [ubiquitin-carrier protein E2]-L-cysteine + [cytochrome c]-N6-ubiquitinyl-L-lysine
-
?
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [histone H3]-L-lysine
-
Oryctolagus cuniculus [ubiquitin-carrier protein E2]-L-cysteine + [histone H3]-N6-ubiquitinyl-L-lysine
-
?
[ubiquitin-carrier protein E2]-S-ubiquitinyl-L-cysteine + [lysozyme]-L-lysine 5% relative activity compared to cytochrome c as substrate Oryctolagus cuniculus [ubiquitin-carrier protein E2]-L-cysteine + [lysozyme]-N6-ubiquitinyl-L-lysine
-
?

Subunits

Subunits Comment Organism
monomer 1 * 230000, SDS-PAGE Oryctolagus cuniculus

Synonyms

Synonyms Comment Organism
arsenite-sensitive E2
-
Oryctolagus cuniculus
E2230K the enzyme prepapration forms a labile addct with a molecular weight of ca. 230 kDa Oryctolagus cuniculus
UB carrier protein (E2)
-
Oryctolagus cuniculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Oryctolagus cuniculus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.003
-
[histone H3]-L-lysine pH 7.0, 37°C, Vmax, 0.06 microM Oryctolagus cuniculus
0.00617
-
[cytochrome c]-L-lysine pH 7.0, 37°C, Vmax, 0.12 microM Oryctolagus cuniculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Oryctolagus cuniculus

General Information

General Information Comment Organism
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