Cloned (Comment) | Organism |
---|---|
- |
Caenorhabditis elegans |
the full-length Sec489Cys mutant as well as the truncated mTR3 missing the C-terminal CUG tripeptide sequence is expressed as a TR-intein-chitin binding domain fusion protein in Escherichia coli | Mus musculus |
wild-type and C-terminal variants | Drosophila melanogaster |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the mutant in the which Sec and Cys residues are switched (TR-GUCG), shows activity similar to that of the Sec489Cys mutant (TR-GCCG). Replacement of the Cys-Sec dyad with a Sec-Sec dyad (TR-GUUG) results in a mutant enzyme with very low catalytic activity. Even if the kcat values are normalized for selenium content, the TR-GUCG and TR-GUUG mutants have catalytic activity 90fold and 185fold lower, respectively, than that of the wild-type enzyme. The mutants in which alanine residues are inserted to increase the ring size (TR-GCAUG and TR-GCAAUG) show only a modest decrease in catalytic activity, 6fold and 4fold, respectively | Mus musculus |
additional information | When the C-terminus of DmTR is changed from a carboxylate to either a thiocarboxylate or a hydroxamic acid, the result is a mutant enzyme with an about 1.7fold increase in activity with thioredoxin. Alanine insertion mutants (DmTR-SCACS and DmTR-SCAACS) show activity with thioredoxin that is greatly reduced compared to that of wild-type DmTR. Increasing the ring size of the Cys-Cys dyad results in a 150-300fold loss in kcat, while the Km is affected little. The 5,5'-dithiobis(2-nitrobenzoic acid) reductase activity of DmTR is also increased when the negative charge at the C-terminus is either neutralized by converting the carboxylate to a neutral hydroxamic acid or modulated by conversion to a thiocarboxylate. Similar to the Sec-containing mammalian enzyme, the truncated DmTR mutant also shows very high 5,5'-dithiobis(2-nitrobenzoic acid) reductase activity, as do the alanine insertion mutants | Drosophila melanogaster |
additional information | where insertion of alanine between the redox-active Cys residues of the C-terminal redox center has very little effect on DTNB reductase activity | Caenorhabditis elegans |
Sec489C | pH-optimum shifts from pH 7.0 to 8.0 | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | KM-values of semisynthetic truncated mTR3 enzymes | Mus musculus | |
additional information | - |
additional information | KM-values of truncated enzymes forms | Drosophila melanogaster | |
0.173 | - |
thioredoxin disulfide | pH 7, 25°C | Drosophila melanogaster | |
0.75 | - |
5,5'-dithiobis(2-nitrobenzoic acid) | pH 7, 25°C | Drosophila melanogaster |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Caenorhabditis elegans | 5739 | - |
mitochondrion | - |
Mus musculus | 5739 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Caenorhabditis elegans | - |
- |
- |
Drosophila melanogaster | - |
- |
- |
Mus musculus | Q8BTW3 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzymes | Caenorhabditis elegans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+ | - |
Drosophila melanogaster | 2-nitro-5-thiobenzoate + NADP+ | - |
? | |
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+ | - |
Caenorhabditis elegans | 2-nitro-5-thiobenzoate + NADP+ | - |
? | |
5,5'-dithiobis(2-nitrobenzoic acid) + NADPH + H+ | C-terminal tetrapeptide sequences is Gly-Cys-Sec-Gly. Changing the C-terminal carboxylate of mTR3 to a carboxamide increases the activity of the enzyme. If the selenium content is normalized for both samples, the carboxamide mutant has nearly twice the activity as the semisynthetic wild-type carboxylate enzyme | Mus musculus | 2-nitro-5-thiobenzoate + NADP+ | - |
? | |
thioredoxin disulfide + NADPH + H+ | - |
Drosophila melanogaster | thioredoxin + NADP+ | - |
? | |
thioredoxin disulfide + NADPH + H+ | - |
Caenorhabditis elegans | thioredoxin + NADP+ | - |
? | |
thioredoxin disulfide + NADPH + H+ | C-terminal tetrapeptide sequences is Gly-Cys-Sec-Gly. Changing the C-terminal carboxylate of mTR3 to a carboxamide increases the activity of the enzyme. If the selenium content is normalized for both samples, the carboxamide mutant has nearly twice the activity as the semisynthetic wild-type carboxylate enzyme | Mus musculus | thioredoxin + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CeTR2 | - |
Caenorhabditis elegans |
DmTR | - |
Drosophila melanogaster |
mTR3 | - |
Mus musculus |
thioredoxin reductase | - |
Drosophila melanogaster |
thioredoxin reductase | - |
Caenorhabditis elegans |
thioredoxin reductase | - |
Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | turnover number of semisynthetic truncated mTR3 enzymes | Mus musculus | |
additional information | - |
additional information | turnover number of truncated enzyme forms | Drosophila melanogaster | |
2.62 | - |
5,5'-dithiobis(2-nitrobenzoic acid) | pH 7, 25°C | Drosophila melanogaster | |
4.99 | - |
thioredoxin disulfide | pH 7, 25°C | Drosophila melanogaster |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
wild-type enzyme | Mus musculus |
7.5 | - |
- |
Drosophila melanogaster |
8 | - |
mutant enzyme Sec489C | Mus musculus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 9 | pH 6.5: about 40% of maximal activity, pH 9.0: about 50% of maximal activity, wild-type enzyme | Mus musculus |
6.5 | 9.5 | pH 6.5: 45% of maximal activity, pH 9.5: about 50% of maximal activity, mutant enzyme Sec489C | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Drosophila melanogaster | |
NADPH | - |
Caenorhabditis elegans | |
NADPH | - |
Mus musculus |