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Literature summary extracted from

  • Stuible, H.; Buttner, D.; Ehlting, J.; Hahlbrock, K.; Kombrink, E.
    Mutational analysis of 4-coumarate:CoA ligase identifies functionally important amino acids and verifies its close relationship to other adenylate-forming enzymes (2000), FEBS Lett., 467, 117-122.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.2.1.12 expressed in Escherichia coli M15[pRP4] as His-tag fusion protein Arabidopsis thaliana

Protein Variants

EC Number Protein Variants Comment Organism
6.2.1.12 C403A activity reduced to 45% of wild-type level, no effect on Km values for ATP and caffeate Arabidopsis thaliana
6.2.1.12 E401Q activity reduced to 21% of wild-type level, no effect on Km values for ATP and caffeate Arabidopsis thaliana
6.2.1.12 K211S activity reduced to 2.7% of wild-type level Arabidopsis thaliana
6.2.1.12 K445T activity reduced to 0.2% of wild-type level Arabidopsis thaliana
6.2.1.12 K457S activity reduced to 4% of wild-type level Arabidopsis thaliana
6.2.1.12 K540N no activity Arabidopsis thaliana
6.2.1.12 R449Q activity reduced to 3% of wild-type level, Km values significantly increased Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.2.1.12 0.005
-
caffeate K457S mutant enzyme Arabidopsis thaliana
6.2.1.12 0.014
-
caffeate K211S mutant enzyme Arabidopsis thaliana
6.2.1.12 0.018
-
caffeate C403A mutant enzyme Arabidopsis thaliana
6.2.1.12 0.024
-
caffeate wild type Arabidopsis thaliana
6.2.1.12 0.028
-
caffeate E401Q mutant enzyme Arabidopsis thaliana
6.2.1.12 0.029
-
ATP K457S mutant enzyme Arabidopsis thaliana
6.2.1.12 0.151
-
ATP K211S mutant enzyme Arabidopsis thaliana
6.2.1.12 0.163
-
ATP wild type Arabidopsis thaliana
6.2.1.12 0.173
-
ATP C403 mutant enzyme Arabidopsis thaliana
6.2.1.12 0.24
-
ATP E401Q mutant enzyme Arabidopsis thaliana
6.2.1.12 0.4
-
caffeate R449Q mutant enzyme Arabidopsis thaliana
6.2.1.12 1.507
-
ATP R449Q mutant enzyme Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.1.12 ATP + 4-coumarate + CoA Arabidopsis thaliana plant phenylpropanoid metabolism AMP + diphosphate + 4-coumaroyl-CoA
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.2.1.12 Arabidopsis thaliana
-
isoform 4CL2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.1.12 recombinant enzyme using His-tag Arabidopsis thaliana

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.2.1.12 0.34
-
K211S mutant enzyme with caffeate as substrate Arabidopsis thaliana
6.2.1.12 0.37
-
R449Q mutant enzyme with caffeate as substrate Arabidopsis thaliana
6.2.1.12 0.46
-
K457S mutant enzyme with caffeate as substrate Arabidopsis thaliana
6.2.1.12 2.64
-
E401Q mutant enzyme with caffeate as substrate Arabidopsis thaliana
6.2.1.12 5.64
-
C403A mutant enzyme with caffeate as substrate Arabidopsis thaliana
6.2.1.12 12.5
-
wild type with caffeate as substrate Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.1.12 ATP + 4-coumarate + CoA
-
Arabidopsis thaliana AMP + diphosphate + 4-coumaroyl-CoA
-
?
6.2.1.12 ATP + 4-coumarate + CoA plant phenylpropanoid metabolism Arabidopsis thaliana AMP + diphosphate + 4-coumaroyl-CoA
-
?
6.2.1.12 ATP + caffeate + CoA
-
Arabidopsis thaliana AMP + diphosphate + caffeoyl-CoA
-
?