Literature summary for 1.8.1.B1 extracted from

Huang, H.H.; Day, L.; Cass, C.L.; Ballou, D.P.; Williams, C.H.; Williams, D.L.
Investigations of the catalytic mechanism of thioredoxin glutathione reductase from Schistosoma mansoni (2011), Biochemistry, 50, 5870-5882.

Application

Application	Comment	Organism
medicine	the enzyme is a drug target for schistosomiasis	Schistosoma mansoni

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Schistosoma mansoni

Protein Variants

Protein Variants	Comment	Organism
C28A	compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 4% activity with GSSG, and 74% activity with thioredoxin disulfide	Schistosoma mansoni
C28A/C31A	compared to the wild type enzyme, the mutant shows 50% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 3% activity with GSSG, and 79% activity with thioredoxin disulfide	Schistosoma mansoni
C31A	compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 18% activity with GSSG, and 100% activity with thioredoxin disulfide	Schistosoma mansoni
C31S	compared to the wild type enzyme, the mutant shows 100% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 129% activity with GSSG, and 105% activity with thioredoxin disulfide	Schistosoma mansoni
C520A	compared to the wild type enzyme, the mutant shows 31% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 26% activity with GSSG, and 47% activity with thioredoxin disulfide	Schistosoma mansoni
C520A/C574A	compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 26% activity with GSSG, and 42% activity with thioredoxin disulfide	Schistosoma mansoni
C574A	compared to the wild type enzyme, the mutant shows 38% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 37% activity with GSSG, and 42% activity with thioredoxin disulfide	Schistosoma mansoni
U597C	compared to the wild type enzyme, the mutant shows 25% activity with 5,5'-dithiobis(2-nitrobenzoic acid), 21% activity with GSSG, and 18% activity with thioredoxin disulfide	Schistosoma mansoni

Inhibitors

Inhibitors	Comment	Organism
auranofin	-	Schistosoma mansoni
furoxan	-	Schistosoma mansoni
potassium antimony tartrate	-	Schistosoma mansoni

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.007	-	thioredoxin disulfide	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.007	-	thioredoxin disulfide	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.008	-	thioredoxin disulfide	mutant enzyme C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.008	-	thioredoxin disulfide	mutant enzyme C31S, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.008	-	thioredoxin disulfide	wild type enzyme, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.013	-	GSSG	mutant enzyme C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.015	-	thioredoxin disulfide	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.015	-	thioredoxin disulfide	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.016	-	GSSG	mutant enzyme C28A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.019	-	thioredoxin disulfide	mutant enzyme C28A/C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.022	-	thioredoxin disulfide	mutant enzyme C28A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.023	-	GSSG	mutant enzyme C28A/C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.024	-	GSSG	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.031	-	GSSG	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.036	-	GSSG	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.042	-	GSSG	wild type enzyme, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.043	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.071	-	GSSG	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.107	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.169	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C31S, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.187	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.189	-	GSSG	mutant enzyme C31S, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.23	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C28A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.23	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.319	-	5,5'-dithiobis(2-nitrobenzoic acid)	wild type enzyme, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.375	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.406	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C28A/C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
GSSG + NADPH + H+	Schistosoma mansoni	-	GSH + NADP+	-	?
thioredoxin disulfide + NADPH + H+	Schistosoma mansoni	-	thioredoxin + NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Schistosoma mansoni	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
2',5'-ADP Sepharose column chromatography and Sephadex G-25 gel filtration	Schistosoma mansoni

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+	-	Schistosoma mansoni	2-nitro-5-thiobenzoate + NADP+	-	?
GSSG + NADPH + H+	-	Schistosoma mansoni	GSH + NADP+	-	?
thioredoxin disulfide + NADPH + H+	-	Schistosoma mansoni	thioredoxin + NADP+	-	?

Synonyms

Synonyms	Comment	Organism
TGR	-	Schistosoma mansoni

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.6	-	GSSG	mutant enzyme C28A/C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
0.7	-	GSSG	mutant enzyme C28A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
3.5	-	GSSG	mutant enzyme C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
3.5	-	thioredoxin disulfide	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
4	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
4	-	GSSG	mutant enzyme U597C, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
5	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
5	-	GSSG	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
5	-	GSSG	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
6	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C28A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
6	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
6	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
6	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
7	-	GSSG	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
8	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C28A/C31A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
8	-	thioredoxin disulfide	mutant enzyme C520A/C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
8	-	thioredoxin disulfide	mutant enzyme C574A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
9	-	thioredoxin disulfide	mutant enzyme C520A, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
16	-	5,5'-dithiobis(2-nitrobenzoic acid)	mutant enzyme C31S, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
16	-	5,5'-dithiobis(2-nitrobenzoic acid)	wild type enzyme, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
19.4	-	GSSG	wild type enzyme, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni
25	-	GSSG	mutant enzyme C31S, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Schistosoma mansoni
NADPH	-	Schistosoma mansoni

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.0000032	-	wild type enzyme, with 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	auranofin
0.0000054	-	wild type enzyme, with GSSG as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	auranofin
0.000008	-	wild type enzyme, with GSSG as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	potassium antimony tartrate
0.0000083	-	wild type enzyme, with thioredoxin disulfide as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	auranofin
0.000009	-	wild type enzyme, with 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	potassium antimony tartrate
0.000045	-	wild type enzyme, with thioredoxin disulfide as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	potassium antimony tartrate
0.003	-	wild type enzyme, with 5,5'-dithiobis(2-nitrobenzoic acid) as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	furoxan
0.0032	-	wild type enzyme, with GSSG as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	furoxan
0.0054	-	wild type enzyme, with thioredoxin disulfide as substrate, in 0.1 M potassium phosphate buffer and 10 mM EDTA (pH 7.4) at 25°C	Schistosoma mansoni	furoxan

General Information

General Information	Comment	Organism
physiological function	the enzyme plays a crucial role in maintaining redox homeostasis	Schistosoma mansoni