Information on EC 4.6.1.12 - 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase

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

EC NUMBER
COMMENTARY hide
4.6.1.12
-
RECOMMENDED NAME
GeneOntology No.
2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol = 2-C-methyl-D-erythritol 2,4-cyclodiphosphate + CMP
show the reaction diagram
; The enzyme from Escherichia coli requires Mg2+ or Mn2+. Forms part of an alternative nonmevalonate pathway for terpenoid biosynthesis
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
formation of diphosphate
-
-
P-O bond cleavage
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Metabolic pathways
-
-
methylerythritol phosphate pathway I
-
-
methylerythritol phosphate pathway II
-
-
Terpenoid backbone biosynthesis
-
-
isoprenoid biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol CMP-lyase (cyclizing; 2-C-methyl-D-erythritol 2,4-cyclodiphosphate-forming)
The enzyme from Escherichia coli requires Mg2+ or Mn2+. Forms part of an alternative nonmevalonate pathway for terpenoid biosynthesis (for diagram, click here).
CAS REGISTRY NUMBER
COMMENTARY hide
287480-92-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain Rv3582c
-
-
Manually annotated by BRENDA team
strain NF54
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
MR1
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Vitis vinifera x Vitis vinifera
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
2-C-methyl-D-erythritol 2,4-cyclodiphosphate + CMP
show the reaction diagram
2-phospho-4-(cytidine 5'-diphospho)-2C-methyl-D-erythritol
2-phospho-2C-methyl-D-erythritol 3,4-cyclophosphate + CMP
show the reaction diagram
2-phospho-4-(cytidine 5'-diphospho)-2C-methyl-D-erythritol
2C-methyl-D-erythritol-2,4-cyclodiphosphate + CMP
show the reaction diagram
4-diphosphocytidyl-2C-methyl-D-erythritol
2C-methyl-D-erythritol 3,4-cyclophosphate + CMP
show the reaction diagram
at low rate
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
2-C-methyl-D-erythritol 2,4-cyclodiphosphate + CMP
show the reaction diagram
2-phospho-4-(cytidine 5'-diphospho)-2C-methyl-D-erythritol
2C-methyl-D-erythritol-2,4-cyclodiphosphate + CMP
show the reaction diagram
P62368
nonmevalonate pathway of isoprenoid biosynthesis
-
-
?
additional information
?
-
-
depletion of MEC synthase has an early and significant impact on cell wall biosynthesis and leads ultimately to cell death
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
preferred
Na+
-
presence of tetrahedral Zn2+ in one of the metal-binding sites and an octahedral sodium ion in the second metal site in absence of substrate
Zinc
-
tightly binds one zinc ion per subunit of the trimer at the active site, which helps to position the substrate for direct attack of the 2-phosphate group on the beta-phosphate
additional information
-
the tetrahedrally arranged transition metal binding site, potentially occupied by Mn2+, sits at the base of the active site cleft. A phosphate oxygen of 2-C-methyl-D-erythritol-2,4-cyclodiphosphate and the side chains of Asp8, His10, and His42 occupy the metal side chains of Asp8, His10, and His42 occupy the metal side coordination sphere
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(+)-(S)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-(2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-5,7-dimethyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylic acid
(+/-)-(2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylic acid
(+/-)-benzyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-benzyl 6-methyl-4-(2-thienyl)-2-thioxo-1,2,3,4-tetra-hydropyrimidine-5-carboxylate
(+/-)-ethyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-5-(4-methoxyphenyl)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-(3,5-difluoro-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-(3-bromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-(4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-(4-methoxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-2-[4-(acetyloxy)-3,5-dibromobenzylidene]-5-(1-benzofuran-2-yl)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo-[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dichloro-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
(+/-)-ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydro-pyrimidine-5-carboxylate
(-)-(R)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
2-amino-N-hydroxy-3-(1-H-indol-3-yl)-propionimidic acid
-
highest affinity of all the ligands examined, due to Zn2+-hydroxamate coordination, KD value of 0.0019 mM
4-amino-1-(3-O-phosphono-a-D-lyxofuranosyl)pyrimidin-2(1H)-one
-
-
4-amino-1-[(2S,3aS,4S,6R,6aR)-2-hydroxy-6-(hydroxymethyl)-2-oxidotetrahydrofuro[3,4-d][1,3,2]dioxaphosphol-4-yl]pyrimidin-2(1H)-one
-
KD value of 20.5 mM
4-amino-1-[(4aR,6R,7R,7aS)-2,7-dihydroxy-2-oxidotetrahydro-4H-furo[3,2-d][1,3,2]dioxaphosphinin-6-yl]pyrimidin-2(1H)-one
-
KD value of 0.925 mM
5-fluorocytidine
-
KD value of 2.02 mM
Cidofovir
-
KD value of ca. 40 mM
Cytosine arabinoside monophosphate
-
KD value of 18.91 mM
EDTA
5 mM. Activity can be restored by the addition of Mg2+ to a concentration of 10 mM
N-([4-[(4-amino-2-oxopyrimidin-1(2H)-yl)methyl]-1-naphthyl]methyl)-4-chlorobenzamide
-
1 mM, 52% inhibition
N-([4-[(4-amino-2-oxopyrimidin-1(2H)-yl)methyl]-1-naphthyl]methyl)benzamide
-
1 mM, 59% inhibition
N-[4-[(6-aminopyridin-3-yl)amino]-3-methylbenzyl]-4-(trifluoromethyl)benzamide
-
1 mM, 60% inhibition
additional information
-
5-(1,3-benzodioxol-5-ylamino)-5-oxopentanoic acid, 2-[(2,6-diaminopyrimidin-4-yl)sulfanyl]acetamide and 4-(5-amino-1,3,4-oxadiazol-2-yl)-1,2,5-oxadiazol-3-amine are inactive. Activity with compounds 2-amino-N-hydroxy-acetimidic acid, N-hydroxy-nicotinimidic acid, 5-bromo-thiophene-2-sulfonic acid amide, 4-bromobenzenesulfonamide and 4-(4-dimethylamino-phenylazo)-benzenesulfonamide are not measurable
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.41 - 2
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
0.252
4-diphosphocytidyl-2C-methyl-D-erythritol
pH 7.0, 27°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.8 - 2.7
2-phospho-4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0089 - 0.06
(+)-(S)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.02 - 0.192
(+/-)-(2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-5,7-dimethyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylic acid
0.018 - 0.223
(+/-)-(2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylic acid
0.002 - 0.069
(+/-)-benzyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0176 - 0.3
(+/-)-benzyl 6-methyl-4-(2-thienyl)-2-thioxo-1,2,3,4-tetra-hydropyrimidine-5-carboxylate
0.0015 - 0.018
(+/-)-ethyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-5-(4-methoxyphenyl)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0015 - 0.032
(+/-)-ethyl (2Z)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.089 - 0.322
(+/-)-ethyl (2Z)-2-(3,5-difluoro-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0016 - 0.212
(+/-)-ethyl (2Z)-2-(3-bromo-4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0032 - 0.3
(+/-)-ethyl (2Z)-2-(4-hydroxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0028 - 0.3
(+/-)-ethyl (2Z)-2-(4-methoxybenzylidene)-7-methyl-3-oxo-5-(2-thienyl)-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.3
(+/-)-ethyl (2Z)-2-[4-(acetyloxy)-3,5-dibromobenzylidene]-5-(1-benzofuran-2-yl)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo-[3,2-a]pyrimidine-6-carboxylate
0.0039 - 0.058
(+/-)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0019 - 0.036
(+/-)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dichloro-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.0063 - 0.3
(+/-)-ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydro-pyrimidine-5-carboxylate
0.015 - 0.039
(-)-(R)-ethyl (2Z)-5-(1-benzofuran-2-yl)-2-(3,5-dibromo-4-hydroxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate
0.000007
artesunate
Plasmodium falciparum
P62369
inhibition of strains NF54 as measured by red blood cell assay
0.000013
Chloroquine
Plasmodium falciparum
P62369
inhibition of strains NF54 as measured by red blood cell assay
0.54
N-([4-[(4-amino-2-oxopyrimidin-1(2H)-yl)methyl]-1-naphthyl]methyl)-4-chlorobenzamide
Escherichia coli
-
-
0.45
N-([4-[(4-amino-2-oxopyrimidin-1(2H)-yl)methyl]-1-naphthyl]methyl)benzamide
Escherichia coli
-
-
0.49
N-[4-[(6-aminopyridin-3-yl)amino]-3-methylbenzyl]-4-(trifluoromethyl)benzamide
Escherichia coli
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.47
calculated
8.22
sequence analysis
8.3
-
calculated from amino acid sequence
8.4
calculated from amino acid sequence
9
calculated
9.4
-
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
tender bark, highest level
Manually annotated by BRENDA team
-
highest expression
Manually annotated by BRENDA team
extremely low expression in latex
Manually annotated by BRENDA team
-
2fold higher expression in petals than in leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Burkholderia cenocepacia (strain ATCC BAA-245 / DSM 16553 / LMG 16656 / NCTC 13227 / J2315 / CF5610)
Burkholderia cenocepacia (strain ATCC BAA-245 / DSM 16553 / LMG 16656 / NCTC 13227 / J2315 / CF5610)
Burkholderia cenocepacia (strain ATCC BAA-245 / DSM 16553 / LMG 16656 / NCTC 13227 / J2315 / CF5610)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain 1710b)
Burkholderia pseudomallei (strain K96243)
Burkholderia pseudomallei (strain K96243)
Burkholderia pseudomallei (strain K96243)
Burkholderia pseudomallei (strain K96243)
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)
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)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Francisella tularensis subsp. tularensis (strain SCHU S4 / Schu 4)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Shewanella oneidensis (strain MR-1)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17300
-
3 * 17300
18260
-
mass spectrometry
19300
calculated from amino acid sequence after removal of 59 N-terminal amino acids comprising a chloroplast transit peptide
21348
x * 21348, electrospray mass spectrometry
24800
x * 24800, calculated from amino acid sequence
25400
-
x * 25400, calculated from amino acid sequence
26030
calculated from amino acid sequence, including a N-terminal chloroplast transit peptide
26670
-
calculated from amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
complex formation of IspDF with 4-diphosphocytidyl-2C-methyl-D-erythritol kinase is observed in solution
trimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in complex with cytidine 5-monophosphate, at 2.3 A resolution. In contrast to bacterial enzymes, the cavity of Arabidopsis thaliana structure is unsuited for binding a diphosphate moiety
-
hanging drop vapor diffusion method, crystal structure of IspDF, a bifunctional methylerythritol 4-phosphate cytidyltransferase methylerythritol 2,4-cyclodidiphosphate synthase
-
3.1 A resolution crystal structure of the Met142/Leu144 mutant
-
computational model of binding of inhibitor N-[4-[(6-aminopyridin-3-yl)amino]-3-methylbenzyl]-4-(trifluoromethyl)benzamide in the active site
-
hanging-drop vapour diffusion method, X-ray crystal structures refined to 2.8 A resolution. The first structure contains a bound Mn2+ cation and the second structure contains CMP, 2-C-methyl-D-erythritol-2,4-cyclodiphosphate, and Mn2+
-
IspF-CDP complex, to 1.8 A resolution. IspF in complex with compound 6b, to 3.1 A resolution, belongs to space group I213 with unit cell parameter a = 144 A. IspF in complex with compound 7, to 2.7 A resolution, belongs to space group I213 with unit cell parameter a = 144.5 A. IspF in complex with cytosine arabinoside monophosphate, to 2.1 A resolution, belongs to space group P21 with unit cell parameters a = 88.8 A, b = 54.2 A, c = 118.4 A, beta = 95°. IspF in complex with 5-fluorocytidine, to 2.5 A resolution, belongs to space group C2 with unit cell parameters a = 104.68 A, b = 54.83 A, c = 88.51 A, beta = 99.66°. IspF in complex with cidofovir, to 2.8 A resolution, belongs to space group I213 with unit cell parameter a = 145.68 A
-
vapor-diffusion hanging drop method, crystal structure of the zinc enzyme in complex with cytidine 5'-diphosphate and Mn2+ is determined to 1.8 A resolution
-
hanging drop vapor diffusion method
high-resolution structure, 16 A, of the enzyme in absence of substrate in the active site. Optimized crystals are obtained at a protein concentration of 35 mg/ml in a solution containing 4 M sodium formate and 5% glycerol. The crystals grow to avarage dimensions of 0.4 * 0.3 * 0.3 mM within a week
-
crystals grow in space group P4(1)2(1)2 from polyethylene glycol using the hanging-drop method
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by metal chelation chromatography
by metal chelation chromatography, more than 95% pure
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 339 bp core fragment and full-length DNA of MECPS amplified and subcloned into pGEM T-easy vector. Escherichia coli harboring both of the plasmids, pAC-BETA and pTrc or pTrcTmMECPS or pTrcAtMECPS
amplification product digested with BamHI and PstI and ligated into plasmid pQE30, expressed in Escherichia coli strain XL1-Blue
cloned into pET17b for expression of the native polypeptide in Escherichia coli strain BL21
expressed in Escherichia coli
expressed in Escherichia coli strain BL21 (DE3)
-
expressed in Escherichia coli strain EB370
expression in Escherichia coli
expression in Escherichia coli strain NMW26 and Arabidopsis thaliana
final amplification product digested with the restriction endonucleases BamHI and SalI and cloned into plasmid pQE30, expressed in Escherichia coli strain XL1-Blue
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
abscisic acid does not modulate enzyme expression
at 4°C, the enzyme expression is down-regulated at 0.05-0.1 mM indole 3-acetic acid. Methyl jasmonate and kinetin down-regulate the expression at 2 h and onwards
enzyme expression in seedlings treated with abscisic acid, H2O2 and drought is higher than in control seedlings
-
enzyme expression is higher in response to light as compared to plants kept under dark.The enzyme also exhibits a gradual up-regulation up to 6 h in response to gibberellic acid treatment
enzyme expression is up-regulated by exogenous elicitors including 0.1 mM methyl jasmonate, acetyl salicylic acid, absisic acid, and UV light
-
higher expression levels in tender barks and leaves than that in roots and stems
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Arg142Met, Glu144Leu
-
dual mutation with little influence on both the overall structure and the detail in the active site
additional information
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Arabidopsis enzyme null mutants are albino lethal. Chloroplasts of mutants are filled with vesicles instead of thylakoids. Expression of enzyme in Escherichia coli can rescue the lethal phenotype of an Escherichia coli ispF mutant
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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high-throughput method for the screening of 2C-methyl-D-erythritol synthase, IspC protein, 4-diphosphocytidyl-2C-methyl-D-erythritol synthase, IspD protein, 4-diphosphocytidyl-2C-methyl-D-erythritol kinase, IspE protein, and 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase, IspF protein, against large compound libraries
drug development
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framework for the development of IspF inhibitors to generate lead compounds of therapeutic potential against microbial pathogens
medicine
the enzyme is a potential target for antimalarial drugs directed at the nonmevalonate pathway of isoprenoid biosynthesis
synthesis
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preparation of 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate in multigram quantities
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