Information on EC 2.7.7.60 - 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
2.7.7.60
-
RECOMMENDED NAME
GeneOntology No.
2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
The enzyme from Escherichia coli requires Mg2+ or Mn2+. ATP or UTP can replace CTP, but both are less effective. GTP and TTP are not substrates. Forms part of an alternative nonmevalonate pathway for terpenoid biosynthesis
-
-
-
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
cytidyltransferase family. Active substrate site consists of a glycine-rich loop spanning Pro13-Arg20. Three-dimensional structure of enzyme is figured out
-
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
ordered sequential mechanism
-
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
no kinetic evidence for substrate channelling
-
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
third step in MEP pathway, isoprenoid biosynthesis
-
CTP + 2-C-methyl-D-erythritol 4-phosphate = diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
third step in MEP pathway, isoprenoid biosynthesis
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
nucleotidyl group transfer
-
-
-
-
nucleotidyl group transfer
;
nucleotidyl group transfer
-
nucleotidyl group transfer
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
methylerythritol phosphate pathway I
-
-
methylerythritol phosphate pathway II
-
-
isoprenoid biosynthesis
-
-
Terpenoid backbone biosynthesis
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
CTP:2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase
The enzyme from Escherichia coli requires Mg2+ or Mn2+. ATP or UTP can replace CTP, but both are less effective. GTP and TTP are not substrates. Forms part of an alternative nonmevalonate pathway for terpenoid biosynthesis (for diagram, click here).
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2-C-methyl erythritol 4-phosphate cytidylyltransferase
-
-
-
-
2-C-methyl-D-erythritol 4-phosphate cytidyltransferase
-
-
2-C-methyl-D-erythritol 4-phosphate cytidyltransferase
-
-
-
2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase
-
2-C-methyl-D-erythritol-4-phosphate cytidylyltransferase
-
-
-
-
2-C-methylerythritol 4-cytidylyltransferase
-
-
-
-
2C-methyl-D-erythritol-4-phosphate cytidylyltransferase
-
2C-methyl-D-erythritol-4-phosphate cytidylyltransferase
-
-
4-diphosphocytidyl-2-C-methyl-D-erythritol synthetase
-
-
-
-
4-diphosphocytidyl-2-C-methylerythritol synthase
-
-
-
-
4-diphosphocytidyl-2C-methyl-D-erythritol synthase
-
-
-
-
CDP-ME synthetase
-
-
-
-
cytidylyltransferase, 2-C-methylerythritol 4-
-
-
-
-
diphosphocytidyl-methylerythritol synthetase
-
-
IspDF
-
bifunctional enzyme catalyzing non-consecutive reactions in the 1-deoxy-D-xylulose 5-phosphate pathway of isoprenoid precursor biosynthesis
MCT
-
-
-
-
MEP cytidyltransferase
-
-
MEP cytidyltransferase
-
-
-
MEP cytidylyltransferase
-
-
-
-
MEP cytidylyltransferase
-
-
MEP cytidylyltransferase
Francisella tularensis FSC155
-
-
-
MEP cytidylyltransferase
-
MtIspD
-
ispD gene from Mycobacterium tuberculosis H37Rv, Rv3582c
MtIspD
-
ispD gene from Mycobacterium tuberculosis H37Rv, Rv3582c
-
CAS REGISTRY NUMBER
COMMENTARY
251990-59-7
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
enzyme gene ispD is fused to gene ispF to encode a bifunctional enzyme catalyzing synthesis of 4-diphosphocytidyl2-C-methyl D-erythritol and of 2-C-methyl-D-erythritol 2,4-cyclodiphosphate
-
-
Manually annotated by BRENDA team
DH5alpha
-
-
Manually annotated by BRENDA team
Escherichia coli DH5-alpha
DH5alpha
-
-
Manually annotated by BRENDA team
Francisella tularensis FSC155
-
-
-
Manually annotated by BRENDA team
Streptomyces coelicolor CH999
CH999
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
enzyme is part of the MEP pathway of biosynthesis of isoprenoid precursors; enzyme is part of the MEP pathway of biosynthesis of isoprenoid precursors
physiological function
2C-methyl-D-erythritol-4-phosphate cytidylyltransferase is the third enzyme in the MEP pathway for isoprenoid biosynthesis
physiological function
-
2C-methyl-D-erythritol-4-phosphate cytidylyltransferase is the third enzyme in the MEP pathway for isoprenoid biosynthesis
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-C-methyl-D-erythritol 4-phosphate + ATP
diphosphate + 4-(adenine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
13% activity compared to CTP as substrate, 30C, 15 min, pH8
-
?
2-C-methyl-D-erythritol 4-phosphate + CTP
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
30C, 15 min, pH8
coupled to hydrolysis of diphosphate to phosphate followed by complex formation with ammonium molybdate-malachite green (absorbance at 630 nm)
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
r
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
ATP, UTP, GTP, ITP, dTTP, dGTP, dCTP ann dATP tested as substrates besides CTP
-
r
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in he mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Streptomyces coelicolor CH999
-
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Francisella tularensis FSC155
-
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
intermediate in the deoxyxylulose pathway of isoprenoid biosynthesis
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
third step of biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
third step of biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Escherichia coli DH5-alpha
-
-
intermediate in the deoxyxylulose pathway of isoprenoid biosynthesis
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Streptomyces coelicolor CH999
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
additional information
?
-
-
monofunctional enzyme
-
-
-
additional information
?
-
-
lipoate-protein ligase attaches octanoate to the dehydrogenase subunit and sulfur insertion protein LipA then converts octanoate to lipoate. LipA acts on both octanoate and octanoyl-proteins
-
-
-
additional information
?
-
-
GTP, UTP and ITP are no substrates
-
-
-
additional information
?
-
-
monofunctional enzyme, GTP, UTP and ITP are no substrates
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
A9ZN09, A9ZN10
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
P9WKG9
-
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in he mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
essential enzyme in the mevalonate-independent pathway of isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol 4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
P9WKG9
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
-
intermediate in the deoxyxylulose pathway of isoprenoid biosynthesis
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
third step of biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
third step of biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Escherichia coli DH5-alpha
-
-
intermediate in the deoxyxylulose pathway of isoprenoid biosynthesis
?
CTP + 2-C-methyl-D-erythritol-4-phosphate
diphosphate + 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
show the reaction diagram
Streptomyces coelicolor CH999
-
part of mevalonate-indpendent pathway for isoprenoid biosynthesis
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Co2+
-
catalytically active with
Divalent cation
-
metal-dependent enzymatic activity
Mg2+
-
essential for activity
Mg2+
-
absolute requirement of divalent metal ions like Ni2+, Mn2+, Co2+, Ca2+, Cu2+ and Zn2+ for catalytic acitvity, maximum activity with Mg2+
Mg2+
-
catalytically active with
Mg2+
-
preferred cation
Mn2+
-
catalytically active with
additional information
-
Cu2+, Ni2+, Ca2+, Fe2+ or Zn2+ do not serve as cofactor
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
CTP
-
substrate inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.043
2-C-methyl-D-erythritol 4-phosphate
-
-
0.043
2-C-methyl-D-erythritol 4-phosphate
calculated affinity
0.061
2-C-methyl-D-erythritol 4-phosphate
-
pH 7.5, 30C
0.178
2-C-methyl-D-erythritol 4-phosphate
-
pH 8.0, 37C
0.31
2-C-methyl-D-erythritol 4-phosphate
-
R20K mutant enzyme, pH 6.5, 23C
0.37
2-C-methyl-D-erythritol 4-phosphate
-
wild type enzyme, pH 6.5, 23C
0.74
2-C-methyl-D-erythritol 4-phosphate
-
R19K mutant enzyme, pH 6.5, 23C
2.98
2-C-methyl-D-erythritol 4-phosphate
-
R19A mutant enzyme, pH 6.5, 23C
3
2-C-methyl-D-erythritol 4-phosphate
-
K27S mutant enzyme, pH 6.5, 23C
3.28
2-C-methyl-D-erythritol 4-phosphate
-
R20A mutant enzyme, pH 6.5, 23C
15.5
2-C-methyl-D-erythritol 4-phosphate
-
D106E mutant enzyme, pH 6.5, 23C
49
2-C-methyl-D-erythritol 4-phosphate
-
T140V mutant enzyme, pH 6.5, 23C
131.5
2-C-methyl-D-erythritol 4-phosphate
-
R109A mutant enzyme, pH 6.5, 23C
0.00314
2-C-methyl-D-erythritol-4-phosphate
-
-
0.032
2-C-methyl-D-erythritol-4-phosphate
-
-
3.26
2-C-methyl-D-erythritol-4-phosphate
-
-
0.131
4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
-
-
0.5
4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
-
-
0.003
CTP
-
-
0.061
CTP
-
pH 7.5, 30C
0.073
CTP
-
pH 8.0, 37C
0.092
CTP
calculated affinity
0.131
CTP
-
-
0.76
CTP
-
wild type enzyme, pH 6.5, 23C
0.85
CTP
-
R19K mutant enzyme, pH 6.5, 23C
1.1
CTP
-
K27S mutant enzyme, pH 6.5, 23C
1.4
CTP
-
D106E mutant enzyme, pH 6.5, 23C
2
CTP
-
T140V mutant enzyme, pH 6.5, 23C
2.8
CTP
-
R19A mutant enzyme, pH 6.5, 23C
3
CTP
-
R109A mutant enzyme, pH 6.5, 23C
3.7
CTP
-
R20K mutant enzyme, pH 6.5, 23C
6.6
CTP
-
R20A mutant enzyme, pH 6.5, 23C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.4
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
R109A mutant enzyme, pH 6.5, 23C
0.75
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
K27S mutant enzyme, pH 6.5, 23C
0.9
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
D106E mutant enzyme, pH 6.5, 23C
1
2-C-methyl-D-erythritol 4-phosphate
Francisella tularensis
-
pH 8.0, 37C
4
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
T140V mutant enzyme, pH 6.5, 23C
21.4
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
R20K mutant enzyme, pH 6.5, 23C
36.8
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
R19A mutant enzyme, pH 6.5, 23C
47.2
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
R19K mutant enzyme, pH 6.5, 23C
48.4
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
wild type enzyme, pH 6.5, 23C
52.7
2-C-methyl-D-erythritol 4-phosphate
Escherichia coli
-
R20A mutant enzyme, pH 6.5, 23C
1.4
2-C-methyl-D-erythritol-4-phosphate
Streptomyces coelicolor
-
-
0.26
CTP
Escherichia coli
-
R109A mutant enzyme, pH 6.5, 23C
0.8
CTP
Francisella tularensis
-
pH 8.0, 37C
0.93
CTP
Escherichia coli
-
D106E mutant enzyme, pH 6.5, 23C
1.8
CTP
Escherichia coli
-
T140V mutant enzyme, pH 6.5, 23C
2.05
CTP
Escherichia coli
-
K27S mutant enzyme, pH 6.5, 23C
3.4
CTP
Mycobacterium tuberculosis
-
slightly lower than from bacterial and plant sources
6
CTP
Streptomyces coelicolor
-
-
9
CTP
Escherichia coli
-
per subunit
26
CTP
Arabidopsis thaliana
-
per subunit
27.5
CTP
Escherichia coli
-
R19A mutant enzyme, pH 6.5, 23C
40.2
CTP
Escherichia coli
-
R20A mutant enzyme, pH 6.5, 23C
46.4
CTP
Escherichia coli
-
R19K mutant enzyme, pH 6.5, 23C
53.6
CTP
Escherichia coli
-
R20K mutant enzyme, pH 6.5, 23C
54.1
CTP
Escherichia coli
-
wild type enzyme, pH 6.5, 23C
16.8
2-C-methyl-D-erythritol-4-phosphate
Escherichia coli
-
-
additional information
ATP
Mycobacterium tuberculosis
-
13% activity compared to CTP
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5.66
2-C-methyl-D-erythritol 4-phosphate
Francisella tularensis
-
pH 8.0, 37C
2166
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
19.6
CTP
-
wild type enzyme, pH 6.5, 23C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.00335
-
assay with D-1-deoxyxylulose-5-phosphate or 2-C-methyl-D-erythritol-4-phosphate
23
-
pure protein
33
-
cell extract
67
-
pure protein
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
assay at
8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
-
sharp decrease in activity above
80
-
no remaining activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
9.1
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
transcription level of enzyme remains constant for at least one month
Manually annotated by BRENDA team
embryonic root, transcription level of enzyme remains constant for at least one month
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Anaerococcus prevotii (strain ATCC 9321 / DSM 20548 / JCM 6508 / PC1)
Burkholderia thailandensis (strain E264 / ATCC 700388 / DSM 13276 / CIP 106301)
Burkholderia thailandensis (strain E264 / ATCC 700388 / DSM 13276 / CIP 106301)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain NCTC 11168)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Listeria monocytogenes serotype 4b (strain F2365)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25900
-
predicted from cDNA
726274
26110
-
deduced from primary sequence; SDS-polyacrylamide gel electrophoresis and mass spectrometry
692926
50000
-
gel filtration
393244
52000
-
native polyacrylamide gel electrophoresis
692926
250000
-
analytical ultracentrifugation
656263
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 27712, ES-MS
?
-
x * 29000, SDS-PAGE, x * 25472 is predicted from sequence of c DNA
?
-
x * 25737 is predicted from sequence of c DNA
?
-
x * 26000, SDS-PAGE
?
x * 36300, calculated
?
Streptomyces coelicolor CH999
-
x * 27712, ES-MS
-
dimer
-
each subunit contains a globular core domain with an alpha/beta sturcture and and one smaller subdomain; homodimer
dimer
-
2 * 25700; homodimer
dimer
-
2 * 26000, SDS-PAGE; homodimer
dimer
-
2 * 25700, crystal structure analysis
dimer
-
crystal structure analysis
dimer
-
gel filtration
dimer
Escherichia coli DH5-alpha
-
2 * 26000, SDS-PAGE; homodimer
-
dimer
Francisella tularensis FSC155
-
gel filtration
-
hexamer
-
6 * 41700, crystal structure analysis
monomer
-
sedimentation velocity experiments, both wild-type and mutant D152A
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
-
-
phosphoprotein
Francisella tularensis FSC155
-
-
-
proteolytic modification
sequence contains a N-terminal chloroplast transit peptide of 88 amino acids
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method
-
catalytic domain of recombinant enzyme, structure contains a cytidine monophosphate bound in the active site suggesting a mechanism for feedback regulation of enzyme activity
hanging drop vapor diffusion method
-
hanging-drop vapour-diffusion using paoyethylene glycol as precipitant, elongated tetragonal prismatic crystals of 0.1 * 0.1 * 0.5 mm, space group P41212
-
vapor diffusion method
-
vapor diffusion method at 4C in hanging drops, crystals belong to space group C2
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25 - 40
-
native-like tertiary structure, shapes of protein similar at 25C, 30C, and 40C
692926
50 - 80
-
unstable at temperature above 50C, estimated by circular dichroism at 25C to 80C (far-UV and near-UV spectra) and after 10 min equilibration, partly integer secondary structure, occurrence of thermally-induced unfolding correlated with catalytic activity loss
692926
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Source 15 Q and Superdex 200 column
-
metal-chelating Hitrap clumn preloaded with Ni2+, Q-Sepharose anion-exchange chromatography to remove previously added thrombin
-
Ni2+-nitrilotriacetic acid column
-
recombinant enzyme using His-tag
-
recombinant protein using His-tag
-
Sepharose Q FF column, Phenyl Sepharose, Cibacron blue 3GA column, Red Sepharose CL-6B column, Source 15 Q column
-
using metal affinity chromatography
-
at 4C, from bacterial lysate by Ni-NTA metal affinity chromatography (elution with 200 mM imidazole), dialysis, concentration by ultrafiltration, yield of 2.8 mg/l culture
-
Q-Sepharose, Phenyl-Sepharose, Superdex HiLoad
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli M15(pREP4)
-
expressed as His-tag fusion protein in Escherichia coli BL21(DE3)
-
expressed as His-tag fusion protein in Escherichia coli M15
-
expressed in Escherichia coli BL21(DE3)
-
hyperexpression in Escherichia coli XL1-Blue
-
overexpression of several genes that could complement the defect of the mutants
-
expressed in Escherichia coli as a His-tagged fusion protein
-
into the vector pT7Blue for sequencing, a cDNA library is constructed in a PDONR222 vector; into the vector pT7Blue for sequencing, a cDNA library is constructed in a PDONR222 vector
from genomic DNA in pET28a for expression with N-terminal hexa-His tag in Escherichia coli BL21 (DE3) for 12 h at 16C
-
expressed in Escherichia coli BL21(DE3)-RP
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D152A
-
catalytically inactive
D106A
-
no activity in crude cell free extract
D106E
-
strongly reduced but detectable activity
D106N
-
no activity in crude cell free extract
K213S
-
effective turnover rate is ca. 0.4% of the rate of wild type, retains capacity to form product with significantly reduced efficiency
K214S
-
no activity in crude cell free extract
K27A
-
amino acid plays an essential role in catalysis because the effective turnover rate is ca. 0.025% of the rate of wild type enzyme
K27A
-
no activity in crude cell free extract
K27S
-
amino acid plays an essential role in catalysis because the effective turnover rate is ca. 0.025% of the rate of wild type enzyme
R109A
-
strongly reduced but detectable activity
R19A
-
no effect on activity
R19K
-
no effect on activity
R20A
-
no effect on activity
R20K
-
reduced activity
T140V
-
strongly reduced but detectable activity
T165V
-
no activity in crude cell free extract
T141D
-
mutant shows only 25% of wild-type activity
T141E
-
mutant catalytically inactive
T141D
Francisella tularensis FSC155
-
mutant shows only 25% of wild-type activity
-
K27S
-
strongly reduced but detectable activity
additional information
-
mutants have been constructed that contain a defect in the pathway to convert 2-C-methyl-D-erythritol 4-phosphate in the unknown product, several genes are cloned to complement the defect of these blocked mutants
T141E
Francisella tularensis FSC155
-
mutant catalytically inactive
-
additional information
heterologous expression in Escherichia coli lacking corresponding enzyme activity rescues the mutant
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
-
high-throughput methods for the screening of 2C-methyl-D-erythritol synthase IspC protein, 4-diphosphocytidyl-2C-methyl-D-erythritol synthase IspD protein, 4-diphosphocytidyl-2Cmethyl-D-erythritol kinase IspE protein, and 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase IspF protein, against large compound libraries. Assays use up to three auxiliary enzymes monitored at 340 and all robust
drug development
-
due to the absence of the pathway in mammals enzyme provides a potential target for new antibiotics
drug development
-
insights in structure and function of MtIspD
medicine
the structure of CMS of Mycobacterium tuberculosis could be a starting point for structure-based design of new antituberculosis drugs
drug development
-
insights in structure and function of MtIspD
-
medicine
-
the structure of CMS of Mycobacterium tuberculosis could be a starting point for structure-based design of new antituberculosis drugs
-