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Literature summary for 2.7.7.60 extracted from

  • Richard, S.B.; Lillo, A.M.; Tetzlaff, C.N.; Bowman, M.E.; Noel, J.P.; Cane, D.E.
    Kinetic analysis of Escherichia coli 2-C-methyl-D-erythritol-4-phosphate cytidyltransferase, wild type and mutants, reveals roles of active site amino acids (2004), Biochemistry, 43, 12189-12197.
    View publication on PubMed
Search for more literature for 2.7.7.60

Cloned(Commentary)

Cloned (Comment) Organism
expressed as His-tag fusion protein in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
D106A no activity in crude cell free extract Escherichia coli
D106E strongly reduced but detectable activity Escherichia coli
D106N no activity in crude cell free extract Escherichia coli
K214S no activity in crude cell free extract Escherichia coli
K27A no activity in crude cell free extract Escherichia coli
K27S strongly reduced but detectable activity Escherichia coli
R109A strongly reduced but detectable activity Escherichia coli
R19A no effect on activity Escherichia coli
R19K no effect on activity Escherichia coli
R20A no effect on activity Escherichia coli
R20K reduced activity Escherichia coli
T140V strongly reduced but detectable activity Escherichia coli
T165V no activity in crude cell free extract Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
CTP substrate inhibition Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.31
-
 2-C-methyl-D-erythritol 4-phosphate R20K mutant enzyme, pH 6.5, 23°C Escherichia coli
0.37
-
 2-C-methyl-D-erythritol 4-phosphate wild type enzyme, pH 6.5, 23°C Escherichia coli
0.74
-
 2-C-methyl-D-erythritol 4-phosphate R19K mutant enzyme, pH 6.5, 23°C Escherichia coli
0.76
-
 CTP wild type enzyme, pH 6.5, 23°C Escherichia coli
0.85
-
 CTP R19K mutant enzyme, pH 6.5, 23°C Escherichia coli
1.1
-
 CTP K27S mutant enzyme, pH 6.5, 23°C Escherichia coli
1.4
-
 CTP D106E mutant enzyme, pH 6.5, 23°C Escherichia coli
2
-
 CTP T140V mutant enzyme, pH 6.5, 23°C Escherichia coli
2.8
-
 CTP R19A mutant enzyme, pH 6.5, 23°C Escherichia coli
2.98
-
 2-C-methyl-D-erythritol 4-phosphate R19A mutant enzyme, pH 6.5, 23°C Escherichia coli
3
-
 2-C-methyl-D-erythritol 4-phosphate K27S mutant enzyme, pH 6.5, 23°C Escherichia coli
3
-
 CTP R109A mutant enzyme, pH 6.5, 23°C Escherichia coli
3.28
-
 2-C-methyl-D-erythritol 4-phosphate R20A mutant enzyme, pH 6.5, 23°C Escherichia coli
3.7
-
 CTP R20K mutant enzyme, pH 6.5, 23°C Escherichia coli
6.6
-
 CTP R20A mutant enzyme, pH 6.5, 23°C Escherichia coli
15.5
-
 2-C-methyl-D-erythritol 4-phosphate D106E mutant enzyme, pH 6.5, 23°C Escherichia coli
49
-
 2-C-methyl-D-erythritol 4-phosphate T140V mutant enzyme, pH 6.5, 23°C Escherichia coli
131.5
-
 2-C-methyl-D-erythritol 4-phosphate R109A mutant enzyme, pH 6.5, 23°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
CTP + 2-C-methyl-D-erythritol 4-phosphate Escherichia coli essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli Q46893
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein using His-tag Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol ordered sequential mechanism Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CTP + 2-C-methyl-D-erythritol 4-phosphate
-
 Escherichia coli diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
-
 ?
CTP + 2-C-methyl-D-erythritol 4-phosphate essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis Escherichia coli diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
-
 ?

Subunits

Subunits Comment Organism
dimer crystal structure analysis Escherichia coli

Synonyms

Synonyms Comment Organism
IspD
-
 Escherichia coli
YgbP
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.26
-
 CTP R109A mutant enzyme, pH 6.5, 23°C Escherichia coli
0.4
-
 2-C-methyl-D-erythritol 4-phosphate R109A mutant enzyme, pH 6.5, 23°C Escherichia coli
0.75
-
 2-C-methyl-D-erythritol 4-phosphate K27S mutant enzyme, pH 6.5, 23°C Escherichia coli
0.9
-
 2-C-methyl-D-erythritol 4-phosphate D106E mutant enzyme, pH 6.5, 23°C Escherichia coli
0.93
-
 CTP D106E mutant enzyme, pH 6.5, 23°C Escherichia coli
1.8
-
 CTP T140V mutant enzyme, pH 6.5, 23°C Escherichia coli
2.05
-
 CTP K27S mutant enzyme, pH 6.5, 23°C Escherichia coli
4
-
 2-C-methyl-D-erythritol 4-phosphate T140V mutant enzyme, pH 6.5, 23°C Escherichia coli
21.4
-
 2-C-methyl-D-erythritol 4-phosphate R20K mutant enzyme, pH 6.5, 23°C Escherichia coli
27.5
-
 CTP R19A mutant enzyme, pH 6.5, 23°C Escherichia coli
36.8
-
 2-C-methyl-D-erythritol 4-phosphate R19A mutant enzyme, pH 6.5, 23°C Escherichia coli
40.2
-
 CTP R20A mutant enzyme, pH 6.5, 23°C Escherichia coli
46.4
-
 CTP R19K mutant enzyme, pH 6.5, 23°C Escherichia coli
47.2
-
 2-C-methyl-D-erythritol 4-phosphate R19K mutant enzyme, pH 6.5, 23°C Escherichia coli
48.4
-
 2-C-methyl-D-erythritol 4-phosphate wild type enzyme, pH 6.5, 23°C Escherichia coli
52.7
-
 2-C-methyl-D-erythritol 4-phosphate R20A mutant enzyme, pH 6.5, 23°C Escherichia coli
53.6
-
 CTP R20K mutant enzyme, pH 6.5, 23°C Escherichia coli
54.1
-
 CTP wild type enzyme, pH 6.5, 23°C Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
19.6
-
 CTP wild type enzyme, pH 6.5, 23°C Escherichia coli