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Literature summary extracted from

  • Bennett, M.R.; Thompson, M.L.; Shepherd, S.A.; Dunstan, M.S.; Herbert, A.J.; Smith, D.R.M.; Cronin, V.A.; Menon, B.R.K.; Levy, C.; Micklefield, J.
    Structure and biocatalytic scope of coclaurine N-methyltransferase (2018), Angew. Chem. Int. Ed. Engl., 57, 10600-10604 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.140 expression in Escherichia coli Coptis japonica

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.140 in complex with S-adenosyl-L-homocysteine and N-methylheliamine. Molecular modeling of structure with natural substrate coclaurine.Both enantiomers of coclaurine can be accommodated in the CNMT active site Coptis japonica

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.140 E204A 4-6% of wild-type activity Coptis japonica
2.1.1.140 E207A 60-75% of wild-type activity Coptis japonica
2.1.1.140 F332A 0.6% of wild-typ activity with substrate coclaurine Coptis japonica
2.1.1.140 G331A negligible effect on activity Coptis japonica
2.1.1.140 H208A 1% of wild-type activity with substrate heliamine, 4% with substrate coclaurine, respectively Coptis japonica
2.1.1.140 R330A negligible effect on activity Coptis japonica
2.1.1.140 W329A 4% of wild-type activity with substrate heliamine, 0.5% with substrate coclaurine, respectively Coptis japonica
2.1.1.140 Y328A 5% of wild-type activity with substrate heliamine, 10% with substrate coclaurine, respectively Coptis japonica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.140 0.265
-
 (RS)-coclaurine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.311
-
 heliamine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.601
-
 (RS)-coclaurine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 1.033
-
 (RS)-coclaurine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 1.565
-
 heliamine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 1.726
-
 (RS)-coclaurine mutant F332A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 2.096
-
 heliamine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.140 Coptis japonica Q948P7
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.140 S-adenosyl-L-methionine + (RS)-coclaurine
-
 Coptis japonica S-adenosyl-L-homocysteine + (RS)-N-methylcoclaurine
-
 ?
2.1.1.140 S-adenosyl-L-methionine + heliamine
-
 Coptis japonica S-adenosyl-L-homocysteine + N-methylheliamine
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.140 0.01
-
 (RS)-coclaurine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.02
-
 (RS)-coclaurine mutant F332A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.12
-
 (RS)-coclaurine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.14
-
 heliamine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.2
-
 heliamine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.53
-
 (RS)-coclaurine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.6
-
 heliamine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.140 0.01
-
 (RS)-coclaurine mutant F332A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.01
-
 (RS)-coclaurine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.09
-
 heliamine mutant W329A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.09
-
 heliamine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 0.2
-
 (RS)-coclaurine mutant Y328A, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 2
-
 (RS)-coclaurine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica
2.1.1.140 2
-
 heliamine wild-type, pH not specified in the publication, temperature not specified in the publication Coptis japonica