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Literature summary for 6.2.1.45 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
Search for more literature for 6.2.1.45

Application

Application Comment Organism
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)

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

Localization

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

Organism

Organism UniProt Comment Textmining
Mus musculus
-
-
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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

Source Tissue Comment Organism Textmining
liver
-
 Mus musculus
-

Storage Stability

Storage Stability Organism
-70°C, recombinant enzyme, no detectable loss of activity Mus musculus
-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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + ubiquitin + [ubiquitin-activating protein E1]-L-cysteine
-
 Mus musculus AMP + diphosphate + [ubiquitin-activating protein E1]-S-ubiquitinyl-L-cysteine
-
 ?
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

Synonyms Comment Organism
ubiquitin-activating enzyme (E1)
-
 Mus musculus

Temperature Optimum [°C]

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

pH Optimum

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