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Literature summary extracted from

  • Lim, J.; Shin, H.; Chung, K.; Kim, N.; Kim, J.; Jung, H.; Im, D.; Jung, C.
    E2-EPF UCP possesses E3 ubiquitin ligase activity via its cysteine 118 residue (2016), PLoS ONE, 11, e0163710 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.24 recombinant expression of diversely tagged wild-type and mutant UCP enzymes in HEK-293T cells, the activity of the recombinant mutant enzymes varies dependent on the fusion tag, overview Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.3.2.24 C118A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.2.24 C95A site-directed mutagenesis, the mutant shows impaired ubiquination activity Homo sapiens
2.3.2.24 K76R/K100R site-directed mutagenesis, the mutant shows reduced autoubiquitination activity compared to wild-type Homo sapiens
2.3.2.24 additional information construction of UCP knockdown cells using a UCP gene-specific shRNA construct. Although UCPDELTAN deletion mutant harbors catalytic active cysteine residues, it cannot ubiquitinate Hippel-Lindau protein (pVHL) due to the missing N-terminus, which forms part of the binding site for pVHL. The UCPC95A/UCPDELTAN pair degrades pVHL in vivo, whereas the UCPDELTAN/UCPC118A pair cannot degrade pVHL. The activity of the recombinant mutant enzymes varies dependent on the fusion tag, overview Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.2.24 [E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [acceptor protein]-L-lysine Homo sapiens
-
 [E1 ubiquitin-activating enzyme]-L-cysteine + [acceptor protein]-N6-monoubiquitinyl-L-lysine
-
 ?
2.3.2.24 [E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [Hippel-Lindau protein]-L-lysine Homo sapiens
-
 [E1 ubiquitin-activating enzyme]-L-cysteine + [Hippel-Lindau protein]-N6-monoubiquitinyl-L-lysine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.24 Homo sapiens Q9C0C9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.2.24 recombinant tagged wild-type and mutant UCP enzymes by affinity chromatography from HEK-293T cells Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.24 additional information E2-EPF ubiquitin carrier protein (UCP) possesses E3 ubiquitin ligase activity via its cysteine 118 residue. E2-EPF UCP elongates E3-independent polyubiquitin chains on the lysine residues of von Hippel-Lindau protein (pVHL) and its own lysine residues both in vitro and in vivo. The initiation of the ubiquitin reaction depends on not only Lys11 linkage but also the Lys6, Lys48 and Lys63 residues of ubiquitin, which are involved in polyubiquitin chain formation on UCP itself. UCP self-association occurred through the UBC domain, which also contributes to the interaction with pVHL. The polyubiquitin chains appears on the N-terminus of UCP in vivo, which indicates that the N-terminus of UCP contains target lysines for polyubiquitination. The Lys76 residue of UCP is the most critical site for auto-ubiquitination, whereas the polyubiquitin chain formation on pVHL occurrs on all three of its lysines (Lys159, Lys171 and Lys196). Polyubiquitin chain formation requires the coordination of Cys95 and Cys118 between two interacting molecules. The mechanism of the polyubiquitin chain reaction of UCP may involve the transfer of ubiquitin from Cys95 to Cys118 by trans-thiolation, with polyubiquitin chains forming at Cys118 by reversible thioester bonding. The polyubiquitin chains are then moved to the lysine residues of the substrate by irreversible isopeptide bonding. During the elongation of the ubiquitin chain, an active Cys118 residue is required in both parts of UCP, namely, the catalytic enzyme and the substrate Homo sapiens ?
-
-
2.3.2.24 [E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [acceptor protein]-L-lysine
-
 Homo sapiens [E1 ubiquitin-activating enzyme]-L-cysteine + [acceptor protein]-N6-monoubiquitinyl-L-lysine
-
 ?
2.3.2.24 [E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [Hippel-Lindau protein]-L-lysine
-
 Homo sapiens [E1 ubiquitin-activating enzyme]-L-cysteine + [Hippel-Lindau protein]-N6-monoubiquitinyl-L-lysine
-
 ?
2.3.2.24 [E1 ubiquitin-activating enzyme]-S-ubiquitinyl-L-cysteine + [Hippel-Lindau protein]-L-lysine the polyubiquitin chain formation on Hippel-Lindau protein (pVHL) occurrs on all three of its lysines (Lys159, Lys171 and Lys196) Homo sapiens [E1 ubiquitin-activating enzyme]-L-cysteine + [Hippel-Lindau protein]-N6-monoubiquitinyl-L-lysine
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.2.24 E2-EPF ubiquitin carrier protein
-
 Homo sapiens
2.3.2.24 E2-EPF UCP
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.2.24 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.24 7.5
-
 assay at Homo sapiens

General Information

EC Number General Information Comment Organism
2.3.2.24 malfunction a UCP mutant in which Cys118 is changed to alanine (UCPC118A) does not form a polyubiquitin chain but strongly accumulates mono- and di-ubiquitin via auto-ubiquitination Homo sapiens
2.3.2.24 additional information UCP ubiquitinates itself independent of E3 ubiquitin ligase in vitro using mixed lysine linkages. E2-EPF ubiquitin carrier protein (UCP) possesses E3 ubiquitin ligase activity via its cysteine 118 residue. E2-EPF UCP elongates E3-independent polyubiquitin chains on the lysine residues of von Hippel-Lindau protein (pVHL) and its own lysine residues both in vitro and in vivo. The initiation of the ubiquitin reaction depends on not only Lys11 linkage but also the Lys6, Lys48 and Lys63 residues of ubiquitin, which are involved in polyubiquitin chain formation on UCP itself. UCP self-association occurred through the UBC domain, which also contributes to the interaction with pVHL. The polyubiquitin chains appears on the N-terminus of UCP in vivo, which indicates that the N-terminus of UCP contains target lysines for polyubiquitination. The Lys76 residue of UCP is the most critical site for auto-ubiquitination, whereas the polyubiquitin chain formation on pVHL occurrs on all three of its lysines (Lys159, Lys171 and Lys196). Polyubiquitin chain formation requires the coordination of Cys95 and Cys118 between two interacting molecules. The mechanism of the polyubiquitin chain reaction of UCP may involve the transfer of ubiquitin from Cys95 to Cys118 by trans-thiolation, with polyubiquitin chains forming at Cys118 by reversible thioester bonding. The polyubiquitin chains are then moved to the lysine residues of the substrate by irreversible isopeptide bonding. During the elongation of the ubiquitin chain, an active Cys118 residue is required in both parts of UCP, namely, the catalytic enzyme and the substrate. Residue Cys118 plays a key role in the autoubiquitination of UCP. Reaction mechanism, overview Homo sapiens
2.3.2.24 physiological function UCP possesses not only E2 ubiquitin conjugating enzyme activity but also E3 ubiquitin ligase activity, and Cys118 is critical for polyubiquitin chain formation Homo sapiens