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Literature summary extracted from

  • Williams, K.M.; Qie, S.; Atkison, J.H.; Salazar-Arango, S.; Alan Diehl, J.; Olsen, S.K.
    Structural insights into E1 recognition and the ubiquitin-conjugating activity of the E2 enzyme Cdc34 (2019), Nat. Commun., 10, 3296 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.3.2.23 additional information the ubiquitin-like modifier-activating enzyme 1 (Uba1, EC 6.2.1.45) is a multidomain enzyme that serves as the gatekeeper of the ubiquitin (Ub) conjugation cascade by activating Ub in a two-step process involving adenylation and thioester bond formation followed by transfer of Ub to E2s in a process termed E1-E2 thioester transfer or transthiolation. Cdc34 is one of tens of E2s that must function with Uba1 despite significant differences at their predicted UFD-interacting surfaces. Molecular recognition of Cdc34 by Uba1, overview. Analysis with Uba1 from Homo sapiens, Saccharomyces cerevisiae, and Schizosaccharomyces pombe Homo sapiens
2.3.2.23 additional information the ubiquitin-like modifier-activating enzyme 1 (Uba1, EC 6.2.1.45) is a multidomain enzyme that serves as the gatekeeper of the ubiquitin (Ub) conjugation cascade by activating Ub in a two-step process involving adenylation and thioester bond formation followed by transfer of Ub to E2s in a process termed E1-E2 thioester transfer or transthiolation. Cdc34 is one of tens of E2s that must function with Uba1 despite significant differences at their predicted UFD-interacting surfaces. Molecular recognition of Cdc34 by Uba1, overview. Analysis with Uba1 from Homo sapiens, Saccharomyces cerevisiae, and Schizosaccharomyces pombe Saccharomyces cerevisiae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.23 gene cdc34, recombinant His-tagged enzyme expression in Escherichia coli strain BL21 (DE3) Codon Plus Homo sapiens
2.3.2.23 gene cdc34, recombinant His-tagged enzyme expression in Escherichia coli strain BL21 (DE3) Codon Plus Saccharomyces cerevisiae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.2.23 Cdc34 alone and in complex with E1, and a Cdc34-Ub thioester mimetic representing the product of Uba1-Cdc34 Ub transthiolation, sitting drop method, mixing of 200 nl of 8.7 mg/ml Uba1-Cdc34DELTAdist crosslink and 0.116 mM Ub, 1 mM ATP, and 5 mM MgCl2, with 200 nl of mother liquor containing 0.2 M ammonium sulfate, 25% PEG 3,350, 0.1 M Bis-Tris, pH 6.5, 18°C, for the free Cdc34DELTAdist, hanging drop method is used with 10 mg/ml protein and mother liquor containing 0.06 M zinc acetate, 0.108 M sodium cacodylate, 14.4% PEG 8000, and 20% glycerol, 18°C, a 1.3fold excess of Cdc34 inhibitor CC0651 is used as a crystallization aid, X-ray diffraction structure determination and analysis at 1.65-2.07 A resolution, molecular replacement using structure PDB ID 3CMM as the search model, modeling Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
2.3.2.23 additional information generation of a Cdc34 active site Cys to Lys mutant Saccharomyces cerevisiae

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.2.23 CC0651
-
 Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine Homo sapiens
-
 [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine Saccharomyces cerevisiae
-
 [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine Saccharomyces cerevisiae ATCC 204508
-
 [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.23 Homo sapiens P49427
-
-
2.3.2.23 Saccharomyces cerevisiae P14682
-
-
2.3.2.23 Saccharomyces cerevisiae ATCC 204508 P14682
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.2.23 recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) Codon Plus by nickel affinity chromatography Homo sapiens
2.3.2.23 recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) Codon Plus by nickel affinity chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 Homo sapiens [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 Saccharomyces cerevisiae [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine the E2 ubiquitin-conjugating enzyme acquires the activated ubquitin from the E1 ubiquitin-activating enzyme Homo sapiens [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine the E2 ubiquitin-conjugating enzyme acquires the activated ubquitin from the E1 ubiquitin-activating enzyme Saccharomyces cerevisiae [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 Saccharomyces cerevisiae ATCC 204508 [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?
2.3.2.23 S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine the E2 ubiquitin-conjugating enzyme acquires the activated ubquitin from the E1 ubiquitin-activating enzyme Saccharomyces cerevisiae ATCC 204508 [E1 ubiquitin-activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.2.23 CDC34
-
 Homo sapiens
2.3.2.23 CDC34
-
 Saccharomyces cerevisiae
2.3.2.23 E2 enzyme
-
 Homo sapiens
2.3.2.23 E2 enzyme
-
 Saccharomyces cerevisiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.23 7.5
-
 E1-E2 thioester transfer assay at Homo sapiens
2.3.2.23 7.5
-
 E1-E2 thioester transfer assay at Saccharomyces cerevisiae

General Information

EC Number General Information Comment Organism
2.3.2.23 metabolism the ubiquitin-like modifier-activating enzyme 1 (Uba1, EC 6.2.1.45) is a multidomain enzyme that serves as the gatekeeper of the ubiquitin (Ub) conjugation cascade by activating Ub in a two-step process involving adenylation and thioester bond formation followed by transfer of Ub to E2s in a process termed E1-E2 thioester transfer or transthiolation. Cdc34 is one of tens of E2s that must function with Uba1 despite significant differences at their predicted UFD-interacting surfaces. Molecular recognition of Cdc34 by Uba1, overview Homo sapiens
2.3.2.23 metabolism the ubiquitin-like modifier-activating enzyme 1 (Uba1, EC 6.2.1.45) is a multidomain enzyme that serves as the gatekeeper of the ubiquitin (Ub) conjugation cascade by activating Ub in a two-step process involving adenylation and thioester bond formation followed by transfer of Ub to E2s in a process termed E1-E2 thioester transfer or transthiolation. Cdc34 is one of tens of E2s that must function with Uba1 despite significant differences at their predicted UFD-interacting surfaces. Molecular recognition of Cdc34 by Uba1, overview Saccharomyces cerevisiae
2.3.2.23 additional information analyis of the molecular basis by which Cdc34 engages its E1, and the structural mechanisms, by which its unique C-terminal extension functions in Cdc34 activity. Conformational changes in Uba1 and Cdc34 and a unique binding mode are required for transthiolation. The Cdc34-Ub structure reveals contacts between the Cdc34 C-terminal extension and Ub that stabilize Cdc34-Ub in a closed conformation and are critical for Ub discharge Homo sapiens
2.3.2.23 additional information analyis of the molecular basis by which Cdc34 engages its E1, and the structural mechanisms, by which its unique C-terminal extension functions in Cdc34 activity. Conformational changes in Uba1 and Cdc34 and a unique binding mode are required for transthiolation. The Cdc34-Ub structure reveals contacts between the Cdc34 C-terminal extension and Ub that stabilize Cdc34-Ub in a closed conformation and are critical for Ub discharge Saccharomyces cerevisiae
2.3.2.23 physiological function ubiquitin (Ub) signaling requires the sequential interactions and activities of three enzymes, E1, E2, and E3. Cdc34 is an E2 that plays a key role in regulating cell cycle progression and requires unique structural elements to function, molecular mechanisms by which Cdc34 function in cells, overview Homo sapiens
2.3.2.23 physiological function ubiquitin (Ub) signaling requires the sequential interactions and activities of three enzymes, E1, E2, and E3. Cdc34 is an E2 that plays a key role in regulating cell cycle progression and requires unique structural elements to function, molecular mechanisms by which Cdc34 function in cells, overview. An ordered Cdc34 CTDprox extension is involved in Ub discharge, CTDprox/Ub contacts are important for Cdc34 action in cells. Cdc34 CTDprox locks Ub(t) in the closed conformation. The Cdc34 CTDprox extension is involved in Ub discharge Saccharomyces cerevisiae