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Literature summary extracted from

  • Carvalho, A.F.; Pinto, M.P.; Grou, C.P.; Vitorino, R.; Domingues, P.; Yamao, F.; Sa-Miranda, C.; Azevedo, J.E.
    High-yield expression in Escherichia coli and purification ofmouse ubiquitin-activating enzyme E1 (2012), Mol. Biotechnol., 51, 254-261.
    View publication on PubMed

Application

EC Number Application Comment Organism
6.2.1.45 synthesis 5–10 mg of histidine-tagged mouse ubiquitin-activating enzyme E1 can be easily obtained from a 1 l Escherichia coli culture. A low temperature during the protein induction step is critical to obtain an active enzyme Mus musculus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.23 expresion in Escherichia coli Mus musculus
6.2.1.45 expresion in Escherichia coli Mus musculus
6.2.1.45 expressed in Escherichia coli BL21(DE3) using pET28-mE1 plasmid Mus musculus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
6.2.1.45 cytosol
-
 Mus musculus 5829
-
6.2.1.45 soluble
-
 Mus musculus
-
-

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.23 Mus musculus Q9Z1K5 isoform UbcH7
-
6.2.1.45 Mus musculus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.1.45 recombinant protein, HIS-Select nickel affinity chromatography, further steps, ion exchange chromatography, followed by gel filtration Mus musculus

Reaction

EC Number Reaction Comment Organism Reaction ID
6.2.1.45 ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine = AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine in the ATP-dependent activation reaction, ubiquitin is covalently attached to a cysteine residue of the E1 component through a thiol ester bond Mus musculus
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
6.2.1.45 liver
-
 Mus musculus
-

Storage Stability

EC Number Storage Stability Organism
6.2.1.45 -70°C, recombinant enzyme, no detectable loss of activity Mus musculus
6.2.1.45 -70°C, the storage of the recombinant mouse E1 does not result in a detectable denaturation of the enzyme Mus musculus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.23 [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine + [ubiquitin carrier protein UbcH7]-L-cysteine
-
 Mus musculus [ubiquitin-activating protein E1]-L-cysteine + [ubiquitin carrier protein UbcH7]-S-ubiquitinyl-L-cysteine
-
 ?
6.2.1.45 ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine
-
 Mus musculus AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine
-
 ?
6.2.1.45 additional information E1 activity is assesssed by the capacity of the enzyme to form a thiol ester conjugate with ubiquitin in an ATP-dependent process and to transfer this activated ubiquitin molecule to an conjugating enzyme Mus musculus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.2.23 ARIH1
-
 Mus musculus
2.3.2.23 UbcH7
-
 Mus musculus
6.2.1.45 ubiquitin-activating enzyme (E1)
-
 Mus musculus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.2.1.45 37
-
 assay at Mus musculus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.2.1.45 7.2
-
 assay at Mus musculus