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Literature summary for 6.2.1.12 extracted from

  • Stuible, H.P.; Kombrink, E.
    Identification of the substrate specificity-conferring amino acid residues of 4-coumarate:coenzyme A ligase allows the rational design of mutant enzymes with new catalytic properties (2001), J. Biol. Chem., 276, 26893-26897.
    View publication on PubMed
Search for more literature for 6.2.1.12

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli M15(pRP4) Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
K320A new substrate specificity, accepts ferulate as substrate Arabidopsis thaliana
K320L new substrate specificity, accepts ferulate as substrate Arabidopsis thaliana
M293A new substrate specificity, accepts ferulate as substrate Arabidopsis thaliana
M293A/V294L substitution of valine decreases Km for ferulate 2-3fold compared to single methionine substitution mutant Arabidopsis thaliana
M293F no reaction with ferulate, increases Km for caffeate Arabidopsis thaliana
M293P new substrate specificity, accepts ferulate as substrate Arabidopsis thaliana
M293P/V294M substitution of valine decreases Km for ferulate 2-3fold compared to single methionine substitution mutant Arabidopsis thaliana
M293W no reaction with ferulate Arabidopsis thaliana
M293Y no reaction with ferulate, increases Km for caffeate Arabidopsis thaliana
V294L slightly reduced activity with caffeate, no activity with ferulate Arabidopsis thaliana
V294M slightly reduced activity with caffeate, no activity with ferulate Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Km values for all mutant enzymes given Arabidopsis thaliana
0.024
-
 caffeate wild type enzyme Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9S725 isoform 4CL2; isoforms 4CL1, 4CL2 and 4CL3, described experiments performed with isoform 4CL2
-

Purification (Commentary)

Purification (Comment) Organism
-
 Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 4-coumarate + CoA
-
 Arabidopsis thaliana AMP + diphosphate + 4-coumaroyl-CoA
-
 ?
ATP + 4-coumarate + CoA plant phenylpropanoid metabolism Arabidopsis thaliana AMP + diphosphate + 4-coumaroyl-CoA
-
 ?
ATP + caffeate + CoA
-
 Arabidopsis thaliana AMP + diphosphate + caffeoyl-CoA
-
 ?
ATP + ferulate + CoA only 4CL1 and 4CL3, no substrate for 4CL2, some 4CL2 mutants are active with ferulate Arabidopsis thaliana AMP + diphosphate + feruloyl-CoA
-
 ?