Literature summary for 1.8.1.7 extracted from

Sarma, G.N.; Savvides, S.N.; Becker, K.; Schirmer, M.; Schirmer, R.H.; Karplus, P.A.
Glutathione reductase of the malarial parasite Plasmodium falciparum: crystal structure and inhibitor development (2003), J. Mol. Biol., 328, 893-907.

Application

Application	Comment	Organism
pharmacology	enzyme is a target for enzyme inhibitor and antimalarial drug development	Plasmodium falciparum

Cloned(Commentary)

Cloned (Comment)	Organism
-	Homo sapiens
expression of strain K1 wild-type and mutant enzymes in Escherichia coli	Plasmodium falciparum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant strain K1 wild-type enzyme, enzyme is dialyzed against EDTA for 3 h, hanging drop vapour diffusion method, protein solution contains 10 mg/ml protein, mixed with an equal volume of reservoir solution containing 14% w/v PEG monomethylether 550, 70 mM NaCl, 70 mM bicine, pH 8.7-9.0, 20°C, one week, X-ray diffraction structure determination and analysis at 3.0 A resolution, molecular replacement	Plasmodium falciparum

Crystallization (Comment)

Organism

purified recombinant strain K1 wild-type enzyme, enzyme is dialyzed against EDTA for 3 h, hanging drop vapour diffusion method, protein solution contains 10 mg/ml protein, mixed with an equal volume of reservoir solution containing 14% w/v PEG monomethylether 550, 70 mM NaCl, 70 mM bicine, pH 8.7-9.0, 20°C, one week, X-ray diffraction structure determination and analysis at 3.0 A resolution, molecular replacement

Plasmodium falciparum

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of a deletion mutant lacking residues 317-349 forms a dimeric FAD-free apoenzyme	Plasmodium falciparum

Inhibitors

Inhibitors	Comment	Organism
1-methyl-4-(2-methyl-1,3-dioxo-2,3,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-5(1H)-yl)pyridinium	poor inhibitor	Homo sapiens
2-methyl-5-(1-naphthyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
2-methyl-5-(1-naphthyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
2-methyl-5-pyridin-4-yl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
2-methyl-5-pyridin-4-yl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
5-(1-anthryl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
5-(1-anthryl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	poor inhibitor	Plasmodium falciparum
5-(4-chlorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
5-(4-chlorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
5-(4-chlorophenyl)-8-fluoro-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
5-(4-chlorophenyl)-8-fluoro-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
5-(pentafluorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
5-(pentafluorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
8-azido-5-(4-chlorophenyl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Homo sapiens
8-azido-5-(4-chlorophenyl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	-	Plasmodium falciparum
ethyl [5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	-	Homo sapiens
ethyl [5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	-	Plasmodium falciparum
ethyl [5-(3-chlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	-	Homo sapiens
additional information	ethyl [5-(3-chlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate and 1-methyl-4-(2-methyl-1,3-dioxo-2,3,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-5(1H)-yl)pyridinium are no inhibitors for the parasite enzyme, numerous noncompetitive inhibitors bind to the large helices-containing cavity at the dimer interface which therefore is a target for selective drugs	Plasmodium falciparum
[5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetic acid	binds to the large helices-containing cavity at the dimer interface	Homo sapiens
[5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetic acid	-	Plasmodium falciparum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.045	-	glutathione disulfide	chloroquine-sensitive strain D10, pH 6.9, 25°C	Plasmodium falciparum
0.065	-	glutathione disulfide	chloroquine-resistant strain FCR3, pH 6.9, 25°C	Plasmodium falciparum
0.071	-	glutathione disulfide	chloroquine-sensitive strain 3D7, pH 6.9, 25°C	Plasmodium falciparum
0.073	-	glutathione disulfide	chloroquine-sensitive strain S106, pH 6.9, 25°C	Plasmodium falciparum
0.074	-	glutathione disulfide	chloroquine-resistant strain Dd2, pH 6.9, 25°C	Plasmodium falciparum
0.076	-	glutathione disulfide	chloroquine-sensitive strain HB3, pH 6.9, 25°C	Plasmodium falciparum
0.08	-	glutathione disulfide	chloroquine-resistant strain 7G8, pH 6.9, 25°C	Plasmodium falciparum
0.088	-	glutathione disulfide	chloroquine-resistant strain K1, pH 6.9, 25°C	Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
glutathione disulfide + NADPH	Plasmodium falciparum	-	glutathione + NADP+	-	r
glutathione disulfide + NADPH	Homo sapiens	-	glutathione + NADP+	-	r
additional information	Plasmodium falciparum	the antioxidant enzyme is important in the parasite sensitive to oxidative stress	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P00390	-	-
Plasmodium falciparum	-	chloroquine-resistant and chloroquine-sensitive strains, overview, intraerythrocytic parasite	-

Reaction

Reaction	Comment	Organism	Reaction ID
2 glutathione + NADP+ = glutathione disulfide + NADPH + H+	active site structure	Homo sapiens	a
2 glutathione + NADP+ = glutathione disulfide + NADPH + H+	active site structure, Cys39/Cys44 are involved	Plasmodium falciparum	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
erythrocyte	-	Homo sapiens	-
trophozoite	-	Plasmodium falciparum	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.009	-	chloroquine-sensitive strain D10, crude extract	Plasmodium falciparum
0.013	-	chloroquine-resistant strains 7G8 and FCR3, crude extract	Plasmodium falciparum
0.022	-	chloroquine-resistant strain Dd2, crude extract	Plasmodium falciparum
0.023	-	chloroquine-sensitive strain S106, crude extract	Plasmodium falciparum
0.033	-	chloroquine-sensitive strain 3D7, crude extract	Plasmodium falciparum
0.035	-	chloroquine-sensitive strain HB3, crude extract	Plasmodium falciparum
0.047	-	chloroquine-resistant strain K1, crude extract	Plasmodium falciparum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
glutathione disulfide + NADPH	-	Plasmodium falciparum	glutathione + NADP+	-	r
glutathione disulfide + NADPH	-	Homo sapiens	glutathione + NADP+	-	r
additional information	the antioxidant enzyme is important in the parasite sensitive to oxidative stress	Plasmodium falciparum	?	-	?

Subunits

Subunits	Comment	Organism
dimer	determination of shape and electrostatics of the large helices-containing cavity at the dimer interface	Plasmodium falciparum
More	enzyme structure comparison with the human enzyme, active site structure	Plasmodium falciparum
More	human enzyme structure comparison with the enzyme from Plasmodium falciparum, human parasite, active site structure	Homo sapiens

Synonyms

Synonyms	Comment	Organism
glutathione reductase	-	Plasmodium falciparum
glutathione reductase	-	Homo sapiens
GR	-	Plasmodium falciparum
GR	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Plasmodium falciparum
25	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.9	-	assay at	Plasmodium falciparum
6.9	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism
FAD	-	Homo sapiens
FAD	flavoenzyme, interaction with Asp311 and Leu352, binding domain structure	Plasmodium falciparum
NADPH	binding site structure	Plasmodium falciparum
NADPH	binding site structure	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.0004	-	5-(4-chlorophenyl)-8-fluoro-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0005	-	2-methyl-5-(1-naphthyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0006	-	5-(4-chlorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0014	-	8-azido-5-(4-chlorophenyl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0019	-	5-(pentafluorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.0025	-	5-(pentafluorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0025	-	2-methyl-5-pyridin-4-yl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0025	-	5-(1-anthryl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Homo sapiens
0.0054	-	5-(4-chlorophenyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.0059	-	5-(4-chlorophenyl)-8-fluoro-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.0062	-	8-azido-5-(4-chlorophenyl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.0066	-	ethyl [5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	pH 6.9, 25°C	Homo sapiens
0.0072	-	[5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetic acid	pH 6.9, 25°C	Homo sapiens
0.0093	-	ethyl [5-(3-chlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	pH 6.9, 25°C	Homo sapiens
0.011	-	2-methyl-5-(1-naphthyl)-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.013	-	ethyl [5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetate	pH 6.9, 25°C	Plasmodium falciparum
0.014	-	[5-(3,5-dichlorophenyl)-1,3-dioxo-3,5,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-2(1H)-yl]acetic acid	pH 6.9, 25°C	Plasmodium falciparum
0.015	-	2-methyl-5-pyridin-4-yl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.2	-	5-(1-anthryl)-2-methyl-5a,9a-dihydropyrido[3,4-b]quinoxaline-1,3(2H,5H)-dione	pH 6.9, 25°C	Plasmodium falciparum
0.2	-	1-methyl-4-(2-methyl-1,3-dioxo-2,3,5a,9a-tetrahydropyrido[3,4-b]quinoxalin-5(1H)-yl)pyridinium	pH 6.9, 25°C	Homo sapiens