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Information on EC 2.1.1.45 - thymidylate synthase and Organism(s) Escherichia coli and UniProt Accession P0A884
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2.1.1.45 thymidylate synthaseSpecify your search results
Word Map
- 2.1.1.45
- 5-fluorouracil
- thymidine
- colorectal
- dihydrofolate
- antifolate
- methotrexate
- dihydropyrimidine
- pyrimidine
-
fluoropyrimidine
- phosphorylase
- ribonucleotide
- leucovorin
- pemetrexed
-
resect
- fdump
- uracil
- mthfr
-
casey
-
schedule
- polyglutamates
-
non-small
- cisplatin
- folylpolyglutamate
- deoxyuridine
- oxaliplatin
- 5-fluoro-2'-deoxyuridine
-
antimetabolite
- orotate
- irinotecan
- capecitabine
-
5-fu-based
-
ccrf-cem
-
folate-dependent
- gemcitabine
-
folinic
-
formyltransferase
- quinazoline
- glycinamide
- dttp
-
polyglutamylation
- 2'-deoxyuridine
-
tegafur
- medicine
-
methotrexate-resistant
- deoxycytidylate
-
cross-complementing
- drug development
-
transformylase
-
5-fu-induced
- dutpase
- aminopterin
- deoxythymidine
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
thymidylate synthase, thymidylate synthetase, human ts, dtmp synthase, thymidylate synthase a, tmp synthetase, 5,10-methylenetetrahydrofolate:dump c-methyltransferase, y110a7a.4, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
dTMP synthase
-
-
-
-
methylenetetrahydrofolate:dUMP C-methyltransferase
-
-
-
-
thymidylate synthetase
-
-
-
-
TMP synthetase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dehalogenation
-
-
-
-
methylene group transfer
-
-
-
-
reductive methylation
reductive methylation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-, -, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
5,10-methylenetetrahydrofolate:dUMP C-methyltransferase
-
CAS REGISTRY NUMBER
COMMENTARY
9031-61-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
-
-
-
?
(R)-5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
?
5-bromo-2'-deoxyuridine 5'-monophosphate + ?
?
dehalogenation does not require methylenetetrahydrofolate as cosubstrate
-
-
?
methylenetetrahydropteroylglutamate + dUMP
7,8-dihydropteroylglutamate + thymidine phosphate
-
-
-
r
(-)-tetrahydropteroylglutamate + 2'-deoxyuridine 5'-monophosphate
7,8-dihydropteroylglutamate + thymidine phosphate
-
-
-
-
?
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
-
-
-
-
?
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
-
r
5,10-methylenetetrahydrofolate + 2'-fluoro-2'-deoxyuridine 5'-phosphate
7,8-dihydrofolate + ?
-
-
-
-
?
5-bromo-2'-deoxyuridine 5'-monophosphate + ?
?
-
dehalogenation
-
-
?
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
-
-
-
?
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
-
-
-
-
?
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
thymidylate synthase is the third enzyme in the methylene tetrahydrofolate cycle besides dihydrofolate reductase and serine hydroxymethyltransferase, in many organisms, thymidylate synthase is the only de novo source for dTMP required for DNA synthesis
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
r
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
r
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
485357, 485363, 485365, 485369, 485371, 485372, 485374, 485382, 485383, 485388, 485397, 485402, 485406, 485409, 485413, 485416, 485417, 485421, 485425, 705044, 705333, 706772, 711474, 757061
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
485357, 485359, 485363, 485365, 485369, 485371, 485372, 485373, 485374, 485375, 485376, 485380, 485381, 485382, 485383, 485388, 485389, 485397, 485402, 485405, 485406, 485408, 485409, 485413, 485415, 485416, 485417, 485419, 485421, 485422, 485423, 485425, 485430, 485431
-
-
r
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
reverse reaction is at least 630times slower than the forward reaction
-
-
r
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
key function in DNA synthesis, repair and cell division
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
biosynthesis of thymidylic acid, only de novo source of thymidine, crucial for DNA replication in every organism
-
-
?
additional information
?
-
chemical conversion takes place preferentially via a concerted mechanism where the hydride transfer is conjugated to thiol-elimination from the product. The substrate-enzyme covalent bond established between the C6 of the nucleotide substrate and a conserved cysteine residue is of labile character
-
-
?
additional information
?
-
-
nucleotides with large substituents on the 2' position are no substrates
-
-
?
additional information
?
-
-
enzyme catalyzes the exchange of a hydrogen atom between water and position 5 of dUMP, occurrence of reactions other than a methyl-group transfer during the interaction of the enzyme with its 2 substrates
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
7,8-dihydrofolate + dTMP
thymidylate synthase is the third enzyme in the methylene tetrahydrofolate cycle besides dihydrofolate reductase and serine hydroxymethyltransferase, in many organisms, thymidylate synthase is the only de novo source for dTMP required for DNA synthesis
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
-
485357, 485363, 485365, 485369, 485371, 485372, 485374, 485382, 485383, 485388, 485397, 485402, 485406, 485409, 485413, 485416, 485417, 485421, 485425
-
-
?
5,10-methylenetetrahydrofolate + dUMP
dihydrofolate + dTMP
-
key function in DNA synthesis, repair and cell division
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(S)-2-(5(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino)1-oxo-2-isoindolinyl)glutaric acid
BW1843U89, U89, distorts the thymidylate synthase active site
2'-deoxy-2',2'-difluorouridine 5'-dihydrogen
classic inhibitor (no time dependence of inhibition observed)
2'-fluoro-ara-UMP
classic inhibitor (no time dependence of inhibition observed)
(2S)-2-[([4-[(2-amino-6-methyl-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)sulfanyl]phenyl]carbonyl)amino]pentanedioic acid
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,4-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]-pyrimidin-4(3H)-one
-
-
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
-
-
2-amino-6-methyl-5-(2-naphthylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
2-amino-6-methyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
5-fluoro-2'-deoxyuridylic acid
-
-
N-(4-((2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo(3,2-d)pyrimidin-5-yl)methyl)benzoyl)-L-glutamic acid
-
-
N-(4-((2-amino-6-methyl-4-oxo-4,7-dihydro-3H-pyrrolo(2,3-d)pyrimidin-5-yl)thio)benzoyl)-L-glutamic acid
-
-
N-[4-[(2,4-diamino-5-methyl-furo[2,3-d]pyrimidin-6-yl)thio]-benzoyl]-L-glutamic acid
-
-
N-[4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)thio]benzoyl]-L-glutamic acid
-
-
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl]-L-glutamic acid
10-propargyl-5,8-dideazafolate
-
i.e. PDDF. Dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Escherichia coli enzyme
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl]-L-glutamic acid
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl]-L-glutamic acid
-
potent inhibitor of thymidylate synthase
pemetrexed
-
dual inhibitor of thymidylate synthase and dihydrofolate reductase. Comparison with inhibitory effect on human and Toxoplasma gondii enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,3-Dimercaptopropanol
-
absolute requirement for a thiol, synthetase shows greatly dimished activity in absence
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.3
(R)-5,10-methylene-5,6,7,8-tetrahydrofolate
30fold higher than wild-type thymidylate synthase
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00027
(2S)-2-[([4-[(2-amino-6-methyl-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)sulfanyl]phenyl]carbonyl)amino]pentanedioic acid
Escherichia coli
-
-
0.023
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
IC50 above 0.023 mM
0.002
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0023
2-amino-5-[(3,4-dichlorophenyl)sulfanyl]-6-methylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0023
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0012
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]-pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0018
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0023
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0022
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0025
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0014
2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.028
2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.002
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0029
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0017
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0024
2-amino-6-methyl-5-(2-naphthylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.0012
2-amino-6-methyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
Escherichia coli
-
-
0.000069
N-(4-((2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo(3,2-d)pyrimidin-5-yl)methyl)benzoyl)-L-glutamic acid
Escherichia coli
-
potent inhibitor of human TS
0.00027
N-(4-((2-amino-6-methyl-4-oxo-4,7-dihydro-3H-pyrrolo(2,3-d)pyrimidin-5-yl)thio)benzoyl)-L-glutamic acid
Escherichia coli
-
moderate inhibitor of human TS
0.23
N-[4-[(2,4-diamino-5-methyl-furo[2,3-d]pyrimidin-6-yl)thio]-benzoyl]-L-glutamic acid
Escherichia coli
-
at 30°C and pH 7.4
0.000018
N-[4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)thio]benzoyl]-L-glutamic acid
Escherichia coli
-
-
0.000069 - 0.00023
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
0.00004
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl]-L-glutamic acid
Escherichia coli
-
-
0.000069
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
Escherichia coli
-
-
0.00023
N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]benzoyl]-L-glutamic acid
Escherichia coli
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 8
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
the nucleotide binds to the free and singly bound forms of the enzyme with nearly equal affinity over a broad range of temperatures and in multiple buffers. There are small but significant differences in DELTACP for the two binding event, so the active sites are not formally equivalent. There is little-to-no allostery at the level of DELTAGbind. There is minor inter-subunit cooperativity in formation of a ternary complex with the mechanism-based inhibitor, 5F-dUMP, and cofactor
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
70000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
dimer
-
-
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
determination of chemical shifts of the apoenzyme (lig0), the saturated holoenzyme (lig2), and the typically elusive singly bound (lig1) states. The two active sites in TS communicate with one another by using the intervening beta-sheet that also forms the dimer interface. The active sites have minimal communication in the lig0 state, but rather a network of correlated motions involving the two active sites is triggered by the first diligand binding event and amplified upon binding the second. Contacts between the diligand and a right-handed beta-bulge feature of this sheet are likely the triggering events
hanging drop method at room temperature, the complexes of mutant TS with dUMP crystallize in the cubic form, binary and ternary complexes with 5NO2dUMP crystallize in the hexagonal form
mutant K48Q, in complex with dUMP or 5-nitro-dUMP, to 2.2 A and 2.7 A resolution, respectively. Binding of dUMP is not impaired in the mutant
crystals of TS apoprotein grown using hanging drop method of vapor diffusion, cubic Laue group m3 with a = 133 A
-
molecular modeling of complex with inhibitor N-[4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidin-5-yl)sulfanyl]benzoyl]-L-glutamic acid. The 6-methyl moiety of of the inhibitor makes important hydrophobic contacts with Trp109 and also serves to lock the 5-position side chain into favorable, low-energy conformations for thymidlyate synthase binding
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K48Q
K48Q
430fold decrease in kcat value, about 30fold increase in Km value for (R)-5,10-methylene-5,6,7,8-tetrahydrofolate. Decrease in binding affinity for folate-like inhibitors
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
43
-
fairly stable, incubation for 10 min results in loss of 50% enzyme activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, enzyme can be stored frozen for several months without significant loss of activity
-
0°C, enzyme can be maintained in its active form in presence of saccharose or glycerol for several days
-
4°C enzyme can be maintained for at least 4 months with little or no loss in activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
after purification and precipitation with ammonium sulfate, wild-type and K48Q TS are frozen at -80°C
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
Escherichia coli transformed with a high amplifying expression plasmid containing the Escherichia coli thymidylate synthase gene
-
overexpression of the encoded gene by transforming Escherichia coli Chi2913
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
-
inhibition of TS and DHFR enzymes has found clinical utility as antitumor, antimicrobial and antiprotozoal agents
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wahba, A.J.; Friedkin, M.
The enzymatic synthesis of thymidylate. I. Early steps in the purification of thymidylate synthetase of Escherichia coli
J. Biol. Chem.
237
3794-3801
1962
Enterococcus faecalis, Escherichia coli, Escherichia coli B / ATCC 11303
Blakeley, R.L.
The biosythesis of thymidylic acid. IV. Further studies on thymidylate synthetase
J. Biol. Chem.
238
2113-2118
1963
Escherichia coli, Enterococcus faecalis, Homo sapiens
-
Fridland, A.; Langenbach, R.J.; Heidelberger, C.
Purification of thymidylate synthetase from Ehrlich ascites carcinoma cells
J. Biol. Chem.
246
7110-7114
1971
Bos taurus, Gallus gallus, Escherichia coli, Enterococcus faecalis, Homo sapiens, Lacticaseibacillus casei, Mus musculus
Horinishi, H.; Greenberg, D.M.
Purification and properties of thymidylate synthase from calf thymus
Biochim. Biophys. Acta
258
741-752
1972
Escherichia phage T2, Enterobacteria phage T6, Bos taurus, Gallus gallus, Escherichia coli, Enterococcus faecalis, Lacticaseibacillus casei, Mus musculus
Santi, D.V.; Sakai, T.T.
Irreversible inhibition of thymidylate synthetase by 5-formyl-2-deoxyuridylic acid
Biochem. Biophys. Res. Commun.
46
1320-1325
1972
Escherichia coli, Escherichia coli B / ATCC 11303, Lacticaseibacillus casei, Mus musculus
Galivan, J.; Maley, G.F.; Maley, F.
Purification and properties of T2 bacteriophage-induced thymidylate synthetase
Biochemistry
13
2282-2289
1974
Escherichia phage T2, Tequatrovirus T4, Enterobacteria phage T6, Bos taurus, Escherichia coli, Lacticaseibacillus casei, Mus musculus, Rattus norvegicus
Friedkin, M.; Plante, L.T.; Crawford, E.J.; Crumm, M.
Inhibition of thymidylate synthetase and dihydrofolate reductase by naturally occurring oligoglutamate derivatives of folic acid
J. Biol. Chem.
250
5614-5621
1975
Escherichia coli, Escherichia coli B / ATCC 11303
McCuen, R.W.; Sirotnak, F.M.
Thymidylate synthetase from Diplococcus pneumoniae, properties and inhibition by folate analogs
Biochim. Biophys. Acta
384
369-380
1975
Enterococcus faecium, Escherichia coli, Escherichia coli B / ATCC 11303, Gallus gallus, Lacticaseibacillus casei, Streptococcus pneumoniae
Slavik, K.; Rode, W.; Slavikova, V.
Purification of thymidylate synthetase from enzyme-poor sources by affinity chromatography
Biochemistry
15
4222-4227
1976
Bos taurus, Escherichia coli, Escherichia coli B / ATCC 11303, Mus musculus
Al Chalabi, K.; Gutteridge, W.E.
Presence and properties of thymidylate synthase in trypanosomatids
Biochim. Biophys. Acta
481
71-79
1977
Crithidia fasciculata, Strigomonas oncopelti, Escherichia coli, Rattus norvegicus, Trypanosoma brucei, Trypanosoma brucei gambiense, Trypanosoma congolense, Trypanosoma cruzi, Trypanosoma lewisi, Trypanosoma rhodesiense
Danenberg, P.V.
Thymidylate synthetase - a target enzyme in cancer chemotherapy
Biochim. Biophys. Acta
473
73-92
1977
Enterococcus faecalis, Escherichia coli, Escherichia coli B / ATCC 11303, Homo sapiens, Lacticaseibacillus casei, Mus musculus, Streptococcus pneumoniae
Dolnick, B.J.; Cheng, Y.C.
Human thymidylate synthetase. II. Derivatives of pteroylmono- and -polyglutamates as substrates and inhibitors
J. Biol. Chem.
253
3563-3567
1978
Escherichia coli, Escherichia coli B / ATCC 11303, Escherichia coli R2, Homo sapiens, Lacticaseibacillus casei
Haertle, T.; Wohlrab, F.; Guschlbauer, W.
Thymidylate synthetase from Escherichia coli K12. Purification, and dependence of kinetic properties on sugar conformation and size of the 2 substituent
Eur. J. Biochem.
102
223-230
1979
Escherichia coli, Escherichia coli B / ATCC 11303, Escherichia coli EM 20031 K12, Escherichia phage T2, Lacticaseibacillus casei, Streptococcus pneumoniae
Lockshin, A.; Moran, R.G.; Danenberg, P.V.
Thymidylate synthetase purified to homogeneity from human leukemic cells
Proc. Natl. Acad. Sci. USA
76
750-754
1979
Escherichia phage T2, Bos taurus, Escherichia coli, Homo sapiens, Lacticaseibacillus casei
Rode, W.; Scanlon, K.J.; Hynes, J.; Bertino, J.R.
Purification of mammalian tumor (L1210) thymidylate synthetase by affinity chromatography on stable biospecific adsorbent. Stabilization of the enzyme with neutral detergents
J. Biol. Chem.
254
11538-11543
1979
Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Mus musculus
Slavik, K.; Slavikova, V.
Purification of thymidylate synthetase from enzyme-poor sources by affinity chromatography
Methods Enzymol.
66
709-723
1980
Bos taurus, Escherichia coli, Escherichia coli B / ATCC 11303, Lacticaseibacillus casei, Mus musculus
Belfort, M.; Moelleken, A.; Maley, G.F.; Maley, F.
Purification and properties of T4 phage thymidylate synthetase produced by the cloned gene in an amplification vector
J. Biol. Chem.
258
2045-2051
1983
Bacillus subtilis, Escherichia coli, Escherichia coli B / ATCC 11303, Escherichia phage T2, Lacticaseibacillus casei, Rattus norvegicus, Tequatrovirus T4
Pattanakitsakul, S.N.; Ruenwongsa, P.
Characterization of thymidylate synthetase and dihydrofolate reductase from Plasmodium berghei
Int. J. Parasitol.
14
513-520
1984
Crithidia fasciculata, Streptococcus pneumoniae, Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Leishmania major, Plasmodium berghei
Matthews, D.A.; Villafranca, J.E.; Janson, C.A.; Smith, W.W.; Welsh, K.; Freer, S.
Stereochemical mechanism of action for thymidylate synthase based on the X-ray structure of the covalent inhibitory ternary complex with 5-fluoro-2-deoxyuridylate and 5,10-methylenetetrahydrofolate
J. Mol. Biol.
214
937-948
1990
Escherichia coli, Lacticaseibacillus casei
Galova, M.; Koptidesova, D.; Rusznakova, D.; Racay, P.; Kollarova, M.
Characteristics of NADPH-dependent thymidylate synthetase purified from Streptomyces aureofaciens
Arch. Biochem. Biophys.
296
81-87
1992
Bos taurus, Saccharomyces cerevisiae, Crithidia fasciculata, Escherichia coli, Mus musculus, Kitasatospora aureofaciens
Garcia-Fuentes, L.; Reche, P.; Lopez-Mayorga, O.; Santi, D.V.; Gonzalez-Pacanowska, D.; Baron, C.
Thermodynamic analysis of the binding of 5-fluoro-2'-deoxyuridine 5'-monophosphate to thymidylate synthase over a range of temperatures
Eur. J. Biochem.
232
641-645
1995
Escherichia coli, Lacticaseibacillus casei
Maley, F.; Pedersen-Lane, J.; Changchien, L.
Complete restoration of activity to inactive mutants of Escherichia coli thymidylate synthase: Evidence that E. coli thymidylate synthase is a half-the-sites activity enzyme
Biochemistry
34
1469-1474
1995
Escherichia coli, Lacticaseibacillus casei
Voeller, D.M.; Changchien, L.M.; Maley, G.F.; Maley, F.; Takechi, T.; Turner, R.E.; Montfort, W.R.; Allegra, C.J.; Chu, E.
Characterization of a specific interaction between Escherichia coli thymidylate synthase and Escherichia coli thymidylate synthase mRNA
Nucleic Acids Res.
23
869-875
1995
Tequatrovirus T4, Escherichia coli, Enterococcus faecium, Homo sapiens
Cho, S.W.; Lee, J.; Cho, E.H.
Effects of mutation at arginine-218 residue on the reaction of synthetic thymidylate synthase and 5-fluorouracil
Exp. Mol. Med.
28
77-83
1996
Tequatrovirus T4, Escherichia coli, Lacticaseibacillus casei, Plasmodium falciparum
-
Hekmat-Nejad, M.; Rathod, P.K.
Kinetics of Plasmodium flaciparum thymidylate synthase: interactions with high-affinity metabolites of 5-fluoroorotate and D1694
Antimicrob. Agents Chemother.
40
1628-1632
1996
Crithidia fasciculata, Escherichia coli, Homo sapiens, Leishmania major, Mus musculus, Plasmodium berghei, Plasmodium falciparum, Plasmodium yoelii
Spencer, H.T.; Villafranca, J.E.; Appleman, J.R.
Kinetic scheme for thymidylate synthase from Escherichia coli: determination from measurements of ligand binding, primary and secondary isotope effects, and pre-steady-state catalysis
Biochemistry
36
4212-4222
1997
Tequatrovirus T4, Candida albicans, Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Mus musculus
Chiericatti, G.; Santi, D.V.
Aspartate 221 of thymidylate synthase is involved in folate cofactor binding and in catalysis
Biochemistry
37
9038-9042
1998
Escherichia coli, Lacticaseibacillus casei
Liang, P.H.; Anderson, K.S.
Substrate channeling and domain-domain interactions in bifunctional thymidylate synthase-dihydrofolate reductase
Biochemistry
37
12195-12205
1998
Escherichia coli, Leishmania major, Toxoplasma gondii
Stout, T.J.; Schellenberger, U.; Santi, D.V.; Stroud, R.M.
Crystal structures of a unique thermal-stable thymidylate synthase from Bacillus subtilis
Biochemistry
37
14736-14747
1998
Lacticaseibacillus casei, Lactococcus lactis, Bacillus phage phi3T, Escherichia coli (P0A884), Escherichia coli, Bacillus subtilis (P0CI79), Bacillus subtilis
Tong, Y.; Liu-Chen, X.; Ercikan-Abali, E.A.; Capiaux, G.M.; Zhao, S.C.; Banerjee, D.; Bertino, J.R.
Isolation and characterization of Thymitaq (AG337) and 5-fluoro-2-deoxyuridylate-resistant mutants of human thymidylate synthase from ethyl methanesulfonate-exposed human sarcoma HT1080 cells
J. Biol. Chem.
273
11611-11618
1998
Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Cryptococcus neoformans (P0CS13), Cryptococcus neoformans B-3501A (P0CS13)
Fox, K.M.; Maley, F.; Garibian, A.; Changchien, L.M.; Van Roey, P.
Crystal structure of thymidylate synthase A from Bacillus subtilis
Protein Sci.
8
538-544
1999
Bacillus subtilis, Escherichia coli
Mahdavian, E.; Spencer, H.T.; Dunlap, R.B.
Kinetic studies on drug-resistant variants of Escherichia coli thymidylate sythase: Functional effects of amino acid sunstitutions at residue 4
Arch. Biochem. Biophys.
368
257-264
1999
Escherichia coli, Homo sapiens
Steadman, D.J.; Spencer, H.T.; Dunlap, R.B.; Berger, S.H.
Substitution at residue 214 of human thymidylate synthase alters nucleotide binding and isomerization of ligand-protein complexes
Biochemistry
38
5582-5587
1999
Escherichia coli, Homo sapiens, Lacticaseibacillus casei
Changchien, L.M.; Garibian, A.; Frasca, V.; Lobo, A.; Maley, G.F.; Maley, F.
High-Level expression of Escherichia coli and Bacillus subtilis thymidylate synthases
Protein Expr. Purif.
19
265-270
2000
Bacillus subtilis, Escherichia coli
Fantz, C.; Shaw, D.; Jennings, W.; Forsthoefel, A.; Kitchens, M.; Phan, J.; Minor, W.; Lebioda, L.; Berger, F.G.; Spencer, H.T.
Drug-resistant variants of Escherichia coli thymidylate synthase: effects of substitutions at Pro-254
Mol. Pharmacol.
57
359-366
2000
Homo sapiens, Mus musculus, Escherichia coli (P0A884), Escherichia coli
Phan, J.; Mahdavian, E.; Nivens, M.C.; Minor, W.; Berger, S.; Spencer, H.T.; Dunlap, R.B.; Lebioda, L.
Catalytic cysteine of thymidylate synthase is activated upon substrate binding
Biochemistry
39
6969-6978
2000
Homo sapiens, Lacticaseibacillus casei, Escherichia coli (P0A884), Escherichia coli
Golos, B.; Dzik, J.M.; Kazimierczuk, Z.; Ciesla, J.; Zielinski, Z.; Jankowska, J.; Kraszewski, A.; Stawinski, J.; Rode, W.; Shugar, D.
Interaction of thymidylate synthase with the 5'-thiophosphates, 5'-dithiophosphates, 5'-H-phosphonates and 5'-S-thiosulfates of 2'-deoxyuridine, thymidine and 5-fluoro-2'-deoxyuridine
Biol. Chem.
382
1439-1445
2001
Escherichia coli, Homo sapiens, Lacticaseibacillus casei, Mus musculus, Pneumocystis carinii, Rattus norvegicus, Trichinella pseudospiralis, Trichinella spiralis
Saxl, R.L.; Changchien, L.M.; Hardy, L.W.; Maley, F.
Parameters affecting the restoration of activity to inactive mutants of thymidylate synthase via subunit exchange: Further evidence that thymidylate synthase Is a half-of-the-sites activity enzyme
Biochemistry
40
5275-5282
2001
Escherichia coli, Lacticaseibacillus casei, Pneumocystis carinii
Fritz, T.A.; Liu, L.; Finer-Moore, J.S.; Stroud, R.M.
Tryptophan 80 and leucine 143 are critical for the hydride transfer step of thymidylate synthase by controlling active site access
Biochemistry
41
7021-7029
2002
Escherichia coli (P0A884), Escherichia coli
Gangjee, A.; Li, W.; Yang, J.; Kisliuk, R.L.
Design, synthesis, and biological evaluation of classical and nonclassical 2-amino-4-oxo-5-substituted-6-methylpyrrolo[3,2-d]pyrimidines as dual thymidylate synthase and dihydrofolate reductase inhibitors
J. Med. Chem.
51
68-76
2008
Escherichia coli, Homo sapiens, Toxoplasma gondii
Arvizu-Flores, A.A.; Sugich-Miranda, R.; Arreola, R.; Garcia-Orozco, K.D.; Velazquez-Contreras, E.F.; Montfort, W.R.; Maley, F.; Sotelo-Mundo, R.R.
Role of an invariant lysine residue in folate binding on Escherichia coli thymidylate synthase: calorimetric and crystallographic analysis of the K48Q mutant
Int. J. Biochem. Cell Biol.
40
2206-2217
2008
Escherichia coli (P0A884), Escherichia coli
Gangjee, A.; Qiu, Y.; Li, W.; Kisliuk, R.L.
Potent dual thymidylate synthase and dihydrofolate reductase inhibitors: classical and nonclassical 2-amino-4-oxo-5-arylthio-substituted-6-methylthieno[2,3-d]pyrimidine antifolates
J. Med. Chem.
51
5789-5797
2008
Escherichia coli, Toxoplasma gondii, Homo sapiens (P04818), Homo sapiens
Jarmula, A.; Dowiercial, A.; Rode, W.
A molecular modeling study of the interaction of 2-fluoro-substituted analogues of dUMP/FdUMP with thymidylate synthase
Bioorg. Med. Chem. Lett.
18
2701-2708
2008
Escherichia coli (P0A884)
Gangjee, A.; Li, W.; Kisliuk, R.L.; Cody, V.; Pace, J.; Piraino, J.; Makin, J.
Design, synthesis, and X-ray crystal structure of classical and nonclassical 2-amino-4-oxo-5-substituted-6-ethylthieno[2,3-d]pyrimidines as dual thymidylate synthase and dihydrofolate reductase inhibitors and as potential antitumor agents
J. Med. Chem.
52
4892-4902
2009
Escherichia coli, Homo sapiens, Toxoplasma gondii
Kanaan, N.; Marti, S.; Moliner, V.; Kohen, A.
QM/MM study of thymidylate synthase: Enzymatic motions and the temperature dependence of the rate limiting step (dagger)
J. Phys. Chem. A
113
2176-2182
2009
Escherichia coli
Arvizu-Flores, A.; Velazquez-Contreras, E.; Machi, L.; Arreola, R.; Maley, F.; Sotelo-Mundo, R.
Spectroscopic analysis of folate binding to thymidylate synthase active site
Spectrosc. Lett.
42
142-146
2009
Escherichia coli
-
Gangjee, A.; Jain, H.D.; Phan, J.; Guo, X.; Queener, S.F.; Kisliuk, R.L.
2,4-Diamino-5-methyl-6-substituted arylthio-furo[2,3-d]pyrimidines as novel classical and nonclassical antifolates as potential dual thymidylate synthase and dihydrofolate reductase inhibitors
Bioorg. Med. Chem.
18
953-961
2010
Escherichia coli, Homo sapiens, Toxoplasma gondii
Islam, Z.; Strutzenberg, T.S.; Ghosh, A.K.; Kohen, A.
Activation of two sequential H-transfers in the thymidylate synthase catalyzed reaction
ACS Catal.
5
6061-6068
2015
Escherichia coli (P0A884), Escherichia coli
Swiderek, K.; Kohen, A.; Moliner, V.
The influence of active site conformations on the hydride transfer step of the thymidylate synthase reaction mechanism
Phys. Chem. Chem. Phys.
17
30793-30804
2015
Escherichia coli (P0A884)
Sapienza, P.J.; Popov, K.I.; Mowrey, D.D.; Falk, B.T.; Dokholyan, N.V.; Lee, A.L.
Inter-active site communication mediated by the dimer interface beta-sheet in the half-the-sites enzyme, thymidylate synthase
Biochemistry
58
3302-3313
2019
Escherichia coli (P0A884)
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