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Literature summary for 1.8.1.9 extracted from

  • Cheng, Q.; Sandalova, T.; Lindqvist, Y.; Arner, E.S.
    Crystal structure and catalysis of the selenoprotein thioredoxin reductase 1 (2009), J. Biol. Chem., 284, 3998-4008.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment) Organism
in the presence of 10 mM NADP+, hanging drop vapor diffusion method, using 15% polyethylene glycol 3350, 12% ethylene glycol, and 0.1 M HEPES, pH 7.5 Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
Y116F the mutant protein is not soluble Rattus norvegicus
Y116I the mutation decreases sensitivity to inhibition by cisplatin and lowers catalytic efficiency in reduction of thioredoxin compared to the wild type enzyme Rattus norvegicus
Y116I the mutation decreases sensitivity to inhibition by cisplatin, lowers catalytic efficiency in reduction of thioredoxin, and increases turnover using 5-hydroxy-1,4-naphthoquinone (juglone) as substrate compared to the wild type enzyme Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
cisplatin complete inhibition after 1 h at 0.5 mM Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00099
-
thioredoxin disulfide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
0.00106
-
thioredoxin disulfide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
0.00143
-
thioredoxin disulfide wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
0.00227
-
5-hydroxy-1,4-naphthoquinone wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
0.00698
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
0.02374
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
0.0906
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
0.094
-
5,5'-dithiobis(2-nitrobenzoic acid) wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
0.0971
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
2.89
-
Lipoamide wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
3.68
-
Lipoamide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
5.59
-
Lipoamide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
selenium thioredoxin reductase 1 is a selenoprotein, the oxidized enzyme presents a selenenylsulfide motif in trans-configuration, with the selenium atom of selenocysteine-498 positioned beneath the side chain of Tyr-116 Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus O89049
-
-

Purification (Commentary)

Purification (Comment) Organism
2',5'-ADP Sepharose column chromatography and phenylarsine oxide-Sepharose column chroamtography Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+
-
Rattus norvegicus 2-nitro-5-thiobenzoate + NADP+
-
?
5-hydroxy-1,4-naphthoquinone + NADPH + H+
-
Rattus norvegicus ?
-
?
lipoamide + NADPH + H+
-
Rattus norvegicus dihydrolipoamide + NADP+
-
?
thioredoxin + insulin disulfide thioredixin is the native principal substrate of TrxR1 Rattus norvegicus thioredoxin disulfide + insulin
-
?

Synonyms

Synonyms Comment Organism
thioredoxin reductase 1
-
Rattus norvegicus
TrxR1
-
Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2 3.7 Lipoamide wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
24.1
-
5-hydroxy-1,4-naphthoquinone wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
27.6
-
Lipoamide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
31.2
-
Lipoamide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
40.98
-
thioredoxin disulfide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
44.85
-
thioredoxin disulfide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
47.72
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
49.87
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
52.75
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
70.3
-
5,5'-dithiobis(2-nitrobenzoic acid) wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
94.17
-
thioredoxin disulfide wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
174.3
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
FAD contains 6 FAD molecules Rattus norvegicus
NADP+ contains 6 NADP+ molecules Rattus norvegicus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.6
-
Lipoamide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
7.5
-
Lipoamide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
513.3
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
526.7
-
5,5'-dithiobis(2-nitrobenzoic acid) mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
748.3
-
5,5'-dithiobis(2-nitrobenzoic acid) wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
7343
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
7557
-
5-hydroxy-1,4-naphthoquinone mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
10620
-
5-hydroxy-1,4-naphthoquinone wild type enzyme, in TE buffer, at 20°C Rattus norvegicus
41450
-
thioredoxin disulfide mutant enzyme Y116I, in TE buffer, at 20°C Rattus norvegicus
42150
-
thioredoxin disulfide mutant enzyme Y116T, in TE buffer, at 20°C Rattus norvegicus
65940
-
thioredoxin disulfide wild type enzyme, in TE buffer, at 20°C Rattus norvegicus