BRENDA - Enzyme Database
show all sequences of 2.1.2.8

Electrostatic guidance of catalysis by a conserved glutamic acid in Escherichia coli dTMP synthase and bacteriophage T4 dCMP hydroxymethylase

Hardy, L.W.; Graves, K.L.; Nalivaika, E.; Biochemistry 34, 8422-8432 (1995)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
overexpression in Escherichia coli BL21; structural gene 42 encodes enzyme
Escherichia virus T4
Engineering
Amino acid exchange
Commentary
Organism
E60D
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
E60Q
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
Inhibitors
Inhibitors
Commentary
Organism
Structure
5-deazatetrahydrofolate
1 mM, 54% inhibition
Escherichia virus T4
5-fluorodeoxyuridylate
mechanism-based inhibitor
Escherichia virus T4
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28487
-
2 * 28487
Escherichia virus T4
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Escherichia virus T4
-
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
Escherichia virus T4
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia virus T4
-
Escherichia coli BL21 infected with bacteriophage T4
-
Purification (Commentary)
Commentary
Organism
purification of wild-type enzyme and mutants E60Q and E60D
Escherichia virus T4
Reaction
Reaction
Commentary
Organism
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
catalytic mechanism; ordered bi-bi mechanism
Escherichia virus T4
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
-
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
485791
Escherichia virus T4
?
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
role of active site Glu-60 is to accelerate bond cleavage between N5 of tetrahydrofolate and the methylene being transferred
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485791
Escherichia virus T4
r
Subunits
Subunits
Commentary
Organism
homodimer
2 * 28487
Escherichia virus T4
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0008
-
deoxycytidylate
mutant E60Q
Escherichia virus T4
0.007
-
deoxycytidylate
mutant E60D
Escherichia virus T4
12.7
-
deoxycytidylate
wild-type enzyme
Escherichia virus T4
Cofactor
Cofactor
Commentary
Organism
Structure
5,10-methylenetetrahydrofolate
-
Escherichia virus T4
tetrahydrofolate
-
Escherichia virus T4
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.11
-
5-deazatetrahydrofolate
-
Escherichia virus T4
Cloned(Commentary) (protein specific)
Commentary
Organism
overexpression in Escherichia coli BL21; structural gene 42 encodes enzyme
Escherichia virus T4
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
5,10-methylenetetrahydrofolate
-
Escherichia virus T4
tetrahydrofolate
-
Escherichia virus T4
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E60D
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
E60Q
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
5-deazatetrahydrofolate
1 mM, 54% inhibition
Escherichia virus T4
5-fluorodeoxyuridylate
mechanism-based inhibitor
Escherichia virus T4
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.11
-
5-deazatetrahydrofolate
-
Escherichia virus T4
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28487
-
2 * 28487
Escherichia virus T4
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Escherichia virus T4
-
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
Escherichia virus T4
?
Purification (Commentary) (protein specific)
Commentary
Organism
purification of wild-type enzyme and mutants E60Q and E60D
Escherichia virus T4
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
-
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
485791
Escherichia virus T4
?
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
role of active site Glu-60 is to accelerate bond cleavage between N5 of tetrahydrofolate and the methylene being transferred
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485791
Escherichia virus T4
r
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 28487
Escherichia virus T4
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0008
-
deoxycytidylate
mutant E60Q
Escherichia virus T4
0.007
-
deoxycytidylate
mutant E60D
Escherichia virus T4
12.7
-
deoxycytidylate
wild-type enzyme
Escherichia virus T4
Other publictions for EC 2.1.2.8
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
735967
Chen
-
Evidence from 18O feeding stud ...
T4virus
Chin. Sci. Bull.
58
864-868
2013
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1
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1
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3
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1
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1
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1
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3
-
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-
659367
Shen
Escherichia coli nucleoside di ...
Escherichia coli
J. Biol. Chem.
279
32225-32232
2004
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1
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3
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1
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1
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1
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-
485792
Song
Crystal structure of deoxycyti ...
Escherichia virus T4
EMBO J.
18
1104-1113
1999
-
-
1
1
1
-
-
-
-
-
1
5
-
2
-
-
1
1
-
-
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-
6
1
-
-
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-
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-
2
-
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1
2
1
1
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1
5
-
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1
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6
1
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-
-
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-
-
-
-
-
485791
Hardy
Electrostatic guidance of cata ...
Escherichia virus T4
Biochemistry
34
8422-8432
1995
-
-
1
-
2
-
2
-
-
-
1
1
-
3
-
-
1
1
-
-
-
-
2
1
-
-
-
3
-
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-
2
1
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1
2
-
2
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2
1
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1
1
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1
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-
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-
2
1
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-
3
-
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-
485789
Graves
Kinetic and equilibrium alpha- ...
Escherichia virus T4
Biochemistry
33
13049-13056
1994
-
-
1
-
3
-
1
-
-
-
1
1
-
2
-
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-
1
-
-
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2
1
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2
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-
1
2
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3
-
-
1
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1
1
-
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-
-
-
-
-
2
1
-
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-
-
-
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-
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-
485790
Butler
Evidence from 18O exchange stu ...
Escherichia virus T4
Biochemistry
33
10521-10526
1994
-
-
1
-
2
-
4
-
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-
1
-
2
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1
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2
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3
1
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2
3
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1
2
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2
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4
3
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1
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2
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3
1
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485788
Mathews
Enzyme interactions involving ...
Escherichia virus T4
Adv. Exp. Med. Biol.
338
563-570
1993
-
-
1
-
1
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2
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1
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3
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2
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1
2
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1
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3
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485787
Subramaniam
On the inhibition of deoxycyti ...
Escherichia virus T4
Arch. Biochem. Biophys.
275
11-15
1989
-
-
-
-
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1
1
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-
1
-
1
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1
1
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1
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2
-
1
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2
1
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2
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1
1
1
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1
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1
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1
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2
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1
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485783
Lamm
Deoxycytidylate hydroxymethyla ...
Enterobacteria phage T2, Enterobacteria phage T6, Escherichia virus T4
Eur. J. Biochem.
172
553-563
1988
-
-
3
-
1
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4
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5
-
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2
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1
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3
1
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1
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6
-
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3
6
-
1
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4
-
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2
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-
1
-
3
1
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1
-
-
-
-
-
-
-
-
-
-
485785
Lee
Deoxycytidylate hydroxymethyla ...
Escherichia virus T4
Biochemistry
27
1367-1373
1988
1
-
-
-
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-
5
1
-
-
2
-
-
2
-
-
1
-
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1
2
1
1
1
-
1
-
1
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2
3
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1
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2
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5
3
1
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2
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1
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1
2
1
1
1
-
1
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1
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485782
Lamm
Nucleotide sequence of the deo ...
Escherichia virus T4
Nucleic Acids Res.
15
3920
1987
-
-
1
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1
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2
-
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1
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2
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1
2
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1
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1
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209854
Allen
T4 phage deoxyribonucleotide-s ...
Escherichia virus T4
J. Biol. Chem.
258
5746-5753
1983
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-
1
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2
2
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1
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3
1
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2
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1
2
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2
2
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3
1
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485786
North
T4 phage-coded deoxycytidylate ...
Escherichia virus T4
Biochem. Biophys. Res. Commun.
77
898-904
1977
-
-
1
-
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1
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3
2
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1
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1
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1
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3
1
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2
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1
2
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1
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3
2
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1
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1
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3
1
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485780
Mathews
Virus-induced acquisition of m ...
bacteriophage T6r+
J. Biol. Chem.
239
2957-2963
1964
-
-
-
-
-
-
-
-
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1
1
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1
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1
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1
1
2
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1
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2
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1
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1
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1
1
2
-
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1
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485781
Pizer
Virus-induced acquisition of m ...
bacteriophage T6r+
J. Biol. Chem.
237
1251-1259
1962
-
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1
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1
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1
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2
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