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Literature summary extracted from

  • Wu, R.; Reger, A.S.; Lu, X.; Gulick, A.M.; Dunaway-Mariano, D.
    The mechanism of domain alternation in the acyl-adenylate forming ligase superfamily member 4-chlorobenzoate: coenzyme A ligase (2009), Biochemistry, 48, 4115-4125.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.2.1.33 mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402 Alcaligenes sp. AL3007

Protein Variants

EC Number Protein Variants Comment Organism
6.2.1.33 D402A 100fold decrease in ratio kcat/Km for CoA Alcaligenes sp. AL3007
6.2.1.33 D402P crystallization data. Mutant adopts the proposed adenylate-forming conformation with very little change to the overall structure.The ability of the mutant to catalyze the adenylate-forming half-reaction is reduced by about 3fold, catalysis of the second half-reaction is reduced by 4 orders of magnitude Alcaligenes sp. AL3007

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.2.1.33 0.0009
-
 4-Chlorobenzoate wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.0012
-
 4-Chlorobenzoate mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.0023
-
 4-Chlorobenzoate mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.0131
-
 ATP mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.0281
-
 ATP mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.124
-
 ATP wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.31
-
 CoA wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 2
-
 CoA mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 6
-
 CoA mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007

Organism

EC Number Organism UniProt Comment Textmining
6.2.1.33 Alcaligenes sp. AL3007
-
-
-
6.2.1.33 Alcaligenes sp. AL3007 Q8GN86 mutant D402P
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.1.33 4-Chlorobenzoate + CoA + ATP
-
 Alcaligenes sp. AL3007 4-Chlorobenzoyl-CoA + AMP + diphosphate
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.2.1.33 0.9
-
 4-Chlorobenzoate mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 1
-
 ATP mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 1.8
-
 CoA mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 2.8
-
 ATP mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 3.4
-
 4-Chlorobenzoate mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 3.7
-
 CoA mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 9.2
-
 4-Chlorobenzoate wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 9.3
-
 CoA wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 11.5
-
 ATP wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.2.1.33 0.02
-
 CoA mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 0.3
-
 CoA mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 2
-
 ATP mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 30
-
 4-Chlorobenzoate mutant D402P, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 30
-
 CoA wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 40
-
 ATP mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 90
-
 ATP wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 400
-
 4-Chlorobenzoate mutant D402A, pH 7.5, 25°C Alcaligenes sp. AL3007
6.2.1.33 10000
-
 4-Chlorobenzoate wild-type, pH 7.5, 25°C Alcaligenes sp. AL3007