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Literature summary for 5.5.1.13 extracted from

  • Koeksal, M.; Potter, K.; Peters, R.J.; Christianson, D.W.
    1.55 A-resolution structure of ent-copalyl diphosphate synthase and exploration of general acid function by site-directed mutagenesis (2014), Biochim. Biophys. Acta, 1840, 184-190.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.5.1.13

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli C41 OverExpress cells Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment) Organism
enzyme complexed with (S)-15-aza-14,15-dihydrogeranylgeranyl thiolodiphosphate, vapor diffusion method, using 100 mM sodium citrate (pH 5.4), 30% (w/v) polyethylene glycol 400, 200 mM KH2PO4 Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
D503A the mutant exhibits a 7fold reduction in kcat Arabidopsis thaliana
E211A the mutation results in a nearly 500fold reduction in kcat Arabidopsis thaliana
N425A the mutant exhibits a 13fold reduction in kcat Arabidopsis thaliana
R340A the mutant exhibits an 850fold reduction in kcat Arabidopsis thaliana
T421A the mutant exhibits a 163fold reduction in kcat (KM is increased 2fold) Arabidopsis thaliana
T421S the mutant exhibits a 3fold reduction in kcat Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0008
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425H, at pH 7.8 and 22°C Arabidopsis thaliana
0.0017
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme E211A, at pH 7.8 and 22°C Arabidopsis thaliana
0.0018
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425A, at pH 7.8 and 22°C Arabidopsis thaliana
0.002
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme R340A, at pH 7.8 and 22°C Arabidopsis thaliana
0.0029
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme D503A, at pH 7.8 and 22°C Arabidopsis thaliana
0.003
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421S, at pH 7.8 and 22°C Arabidopsis thaliana
0.003
-
 (E,E,E)-geranylgeranyl diphosphate wild type enzyme, at pH 7.8 and 22°C Arabidopsis thaliana
0.006
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421A, at pH 7.8 and 22°C Arabidopsis thaliana

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for optimal catalytic activity Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(E,E,E)-geranylgeranyl diphosphate Arabidopsis thaliana
-
 ent-copalyl diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q38802
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA His-Bind resin column chromatography Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(E,E,E)-geranylgeranyl diphosphate
-
 Arabidopsis thaliana ent-copalyl diphosphate
-
 ?
additional information no activity with (S)-15-aza-14,15-dihydrogeranylgeranyl thiolodiphosphate Arabidopsis thaliana ?
-
 ?

Synonyms

Synonyms Comment Organism
CPS
-
 Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0011
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme R340A, at pH 7.8 and 22°C Arabidopsis thaliana
0.0019
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme E211A, at pH 7.8 and 22°C Arabidopsis thaliana
0.006
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421A, at pH 7.8 and 22°C Arabidopsis thaliana
0.012
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425H, at pH 7.8 and 22°C Arabidopsis thaliana
0.071
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425A, at pH 7.8 and 22°C Arabidopsis thaliana
0.13
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme D503A, at pH 7.8 and 22°C Arabidopsis thaliana
0.3
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421S, at pH 7.8 and 22°C Arabidopsis thaliana
0.9
-
 (E,E,E)-geranylgeranyl diphosphate wild type enzyme, at pH 7.8 and 22°C Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.55
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme R340A, at pH 7.8 and 22°C Arabidopsis thaliana
1
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421A, at pH 7.8 and 22°C Arabidopsis thaliana
1.1
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme E211A, at pH 7.8 and 22°C Arabidopsis thaliana
20
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425H, at pH 7.8 and 22°C Arabidopsis thaliana
40
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme N425A, at pH 7.8 and 22°C Arabidopsis thaliana
45
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme D503A, at pH 7.8 and 22°C Arabidopsis thaliana
100
-
 (E,E,E)-geranylgeranyl diphosphate mutant enzyme T421S, at pH 7.8 and 22°C Arabidopsis thaliana
300
-
 (E,E,E)-geranylgeranyl diphosphate wild type enzyme, at pH 7.8 and 22°C Arabidopsis thaliana