Literature summary for 1.5.1.3 extracted from

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 (2009), J. Med. Chem., 52, 4892-4902.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme mutant Q35S/N64S in complex with NADPH and inhibitor N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid, and enzyme mutant Q35K in complex with NADPH and inhibitor N-{4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl}-L-glutamic acid, X-ray diffraction structure determination and analysis at 1.2-1.5 A resolution	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one	-	Toxoplasma gondii
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one	-	Escherichia coli
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one	-	Homo sapiens
2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one	-	Toxoplasma gondii
methotrexate	-	Escherichia coli
methotrexate	-	Homo sapiens
methotrexate	-	Toxoplasma gondii
N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Escherichia coli
N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Homo sapiens
N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Toxoplasma gondii
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Escherichia coli
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Homo sapiens
N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid	-	Toxoplasma gondii
nolatrexed	-	Escherichia coli
nolatrexed	-	Homo sapiens
nolatrexed	-	Toxoplasma gondii
pemetrexed	-	Escherichia coli
pemetrexed	-	Homo sapiens
pemetrexed	-	Toxoplasma gondii
permetrexed	-	Escherichia coli
permetrexed	-	Homo sapiens
permetrexed	-	Toxoplasma gondii
piritrexim	-	Escherichia coli
piritrexim	-	Homo sapiens
piritrexim	-	Toxoplasma gondii
plevitrexed	-	Escherichia coli
plevitrexed	-	Homo sapiens
plevitrexed	-	Toxoplasma gondii
raltitrexed	-	Escherichia coli
raltitrexed	-	Homo sapiens
raltitrexed	-	Toxoplasma gondii
trimethoprim	-	Escherichia coli
trimethoprim	-	Homo sapiens
trimethoprim	-	Toxoplasma gondii
trimetrexate	-	Escherichia coli
trimetrexate	-	Homo sapiens
trimetrexate	-	Toxoplasma gondii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
7,8-dihydrofolate + NADPH + H+	Escherichia coli	-	5,6,7,8-tetrahydrofolate + NADP+	-	?
7,8-dihydrofolate + NADPH + H+	Toxoplasma gondii	-	5,6,7,8-tetrahydrofolate + NADP+	-	?
7,8-dihydrofolate + NADPH + H+	Homo sapiens	-	5,6,7,8-tetrahydrofolate + NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-
Homo sapiens	P00374	-	-
Toxoplasma gondii	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3)	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
7,8-dihydrofolate + NADPH + H+	-	Escherichia coli	5,6,7,8-tetrahydrofolate + NADP+	-	?
7,8-dihydrofolate + NADPH + H+	-	Toxoplasma gondii	5,6,7,8-tetrahydrofolate + NADP+	-	?
7,8-dihydrofolate + NADPH + H+	-	Homo sapiens	5,6,7,8-tetrahydrofolate + NADP+	-	?

Synonyms

Synonyms	Comment	Organism
DHFR	-	Escherichia coli
DHFR	-	Toxoplasma gondii
DHFR	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Escherichia coli
37	-	assay at	Toxoplasma gondii
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Escherichia coli
7	-	assay at	Toxoplasma gondii
7	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism
NADPH	-	Escherichia coli
NADPH	-	Toxoplasma gondii
NADPH	-	Homo sapiens

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.0000000088	-	pH 7.0, 37°C	Escherichia coli	methotrexate
0.00000001	-	pH 7.0, 37°C	Escherichia coli	trimethoprim
0.000000019	-	pH 7.0, 37°C	Homo sapiens	N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.00000002	-	pH 7.0, 37°C	Homo sapiens	methotrexate
0.00000002	-	pH 7.0, 37°C	Homo sapiens	N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.0000002	-	pH 7.0, 37°C	Escherichia coli	N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.00000026	-	pH 7.0, 37°C	Homo sapiens	2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
0.00000026	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.00000028	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one
0.00000029	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.0000003	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
0.0000003	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.00000035	-	pH 7.0, 37°C	Homo sapiens	2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one
0.00000054	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.0000006	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.00000062	-	pH 7.0, 37°C	Homo sapiens	2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one
0.00000084	-	pH 7.0, 37°C	Homo sapiens	2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000001	-	pH 7.0, 37°C	Escherichia coli	N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.0000021	-	pH 7.0, 37°C	Toxoplasma gondii	N-(4-[(2-amino-6-ethyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.0000022	-	pH 7.0, 37°C	Homo sapiens	2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one
0.0000026	-	pH 7.0, 37°C	Homo sapiens	2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
0.0000028	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one
0.000003	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.0000066	-	pH 7.0, 37°C	Homo sapiens	pemetrexed
0.000008	-	pH 7.0, 37°C	Toxoplasma gondii	N-(4-[(2-amino-6-methyl-4-oxo-3,4-dihydrothieno[2,3-d ]pyrimidin-5-yl)thio]benzoyl)-L-glutamic acid
0.0000081	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.0000084	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one
0.000009	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
0.000012	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(3-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000013	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000014	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000014	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(3,5-dichlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]-pyrimidin-4(3H)-one
0.000017	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one
0.000022	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(3,4-dichlorophenyl)thio]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000024	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000025	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one
0.000027	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(3,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000027	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000028	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(2,5-dimethoxyphenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000028	-	pH 7.0, 37°C	Escherichia coli	2-amino-6-ethyl-5-(2-naphthylthio)thieno[2,3-d ]pyrimidin-4(3H)-one
0.000029	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(4-bromophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000031	-	pH 7.0, 37°C	Escherichia coli	2-amino-6-ethyl-5-[(4-fluorophenyl)sulfanyl]thieno[2,3-d ]pyrimidin-4(3H)-one
0.000033	-	pH 7.0, 37°C	Toxoplasma gondii	methotrexate
0.000033	-	pH 7.0, 37°C	Escherichia coli	2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
0.000033	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-6-ethyl-5-(phenylsulfanyl)thieno[2,3-d]pyrimidin-4(3H)-one
0.000033	-	pH 7.0, 37°C	Escherichia coli	2-amino-6-ethyl-5-(pyridin-4-ylsulfanyl)thieno[2,3-d ]pyrimidin-4(3H)-one
0.000033	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(2-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d ]pyrimidin-4(3H)-one
0.000039	-	pH 7.0, 37°C	Escherichia coli	2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
0.000042	-	pH 7.0, 37°C	Escherichia coli	2-amino-5-[(4-chlorophenyl)sulfanyl]-6-ethylthieno[2,3-d]pyrimidin-4(3H)-one
0.000087	-	pH 7.0, 37°C	Toxoplasma gondii	2-amino-6-ethyl-5-[(4-nitrophenyl)sulfanyl]thieno[2,3-d]pyrimidin-4(3H)-one
0.00023	-	pH 7.0, 37°C	Escherichia coli	pemetrexed
0.00034	-	pH 7.0, 37°C	Homo sapiens	trimethoprim
0.00043	-	pH 7.0, 37°C	Toxoplasma gondii	pemetrexed
0.0068	-	pH 7.0, 37°C	Toxoplasma gondii	trimethoprim