Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.2.1.45 extracted from

  • Zheng, M.; Liu, J.; Yang, Z.; Gu, X.; Li, F.; Lou, T.; Ji, C.; Mao, Y.
    Expression, purification and characterization of human ubiquitin-activating enzyme, UBE1 (2010), Mol. Biol. Rep., 37, 1413-1419.
    View publication on PubMed
Search for more literature for 6.2.1.45

Application

Application Comment Organism
synthesis expression of the full length of human UBE1 in Escherichia coli and purification by Ni-NTA superflow sepharose and strep-tactin sepharose which is based on UB-UBE1 high-energy thioester bonded intermediate complex. Purified UBE1 can activate and conjugate UB to ubiquitin-conjugating enzyme E2s Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Homo sapiens
full length of human UBE1 is expressed in Escherichia coli via pET28b vector Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
117000
-
 x * 117000, SDS-PAGE Homo sapiens
118000
-
 SDS-PAGE Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Homo sapiens Q712V1
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme, purification on Ni-NTA superflow sepharose and strep-tactin sepharose which is based on UB-UBE1 high-energy thioester bonded intermediate complex Homo sapiens
recombinant protein Homo sapiens
recombinant protein is purified by Ni-NTA His-Bind Superflow Sepharose and Strep-Tactin Sepahrose Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
ATP + ubiquitin + [E1 ubiquitin-activating enzyme]-L-cysteine = AMP + diphosphate + S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine ATP-dependant formation of C-S thioester bond between the C-terminal glycine of ubiquitin and a sulfhydryl side group of a cysteine residue on the E1 protein Homo sapiens
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + ubiquitin + [6His-ubiquitin-activating enzyme E1]W-8His-Strep-HA Strep, i.e.WSHPQFEK, HA, i.e. YPYDVPDYAS, under non-reducing conditions, the intermediate complex of the thioester formation is not observed without ATP Homo sapiens AMP + diphosphate + [6His-ubiquitin-activating enzyme E1]W-8His-Strep-HA-ubiquitinyl-L-cysteine
-
 ?
ATP + ubiquitin + [ubiquitin-activating protein UBE1]-L-cysteine
-
 Homo sapiens AMP + diphosphate + [ubiquitin-activating protein UBE1]-S-ubiquitinyl-L-cysteine
-
 ?
additional information the thioester formation assay is performed using recombinant proteins expressed in Escherichia coli. The activation of ubiquitin by purified UBE1 is identified in vitro by SDS-PAGE Homo sapiens ?
-
 ?
additional information purified isoform UBE1 can activate and conjugate ubiquitin to ubiquitin-conjugating enzyme E2s. Transfer is restricted to distinct E2 isoforms UB2R2, UBE2W and UBE2NL Homo sapiens ?
-
 ?

Subunits

Subunits Comment Organism
? x * 117000, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
UBE1
-
 Homo sapiens
ubiquitin-activating enzyme, UBE1
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens

General Information

General Information Comment Organism
physiological function plays an important role in the first step of the proteasome pathway to activate ubiquitin. The UBE1 is a two-step intramolecular and ATP-dependent reaction to generate a high-energy E1-thiol-ester-ubiquitin intermediate. The activated ubiquitin are then transferred to ubiquitin-conjugating enzyme E2 Homo sapiens