Crystallization (Comment) | Organism |
---|---|
TbQSOX crystal structure analysis, PDB ID 3QD9 | Trypanosoma brucei |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the wild-type CGAC motif in the thioredoxin domain of enzyme TbQSOX is replaced by the more oxidizing CPHC or more reducing CGPC sequence, resulting in mutants CIPHCII and CIGPCII. Construction of a truncated mutant containing only the thioredoxin domain TbQSOX-TRX, corresponding to residues 20-199 | Trypanosoma brucei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | stopped-flow kinetic measurements | Trypanosoma brucei | |
0.03 | - |
dithiothreitol | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
0.04 | - |
RNAse A | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
0.17 | - |
dithiothreitol | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei | |
0.22 | - |
dithiothreitol | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
0.32 | - |
RNAse A | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
0.36 | - |
RNAse A | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
RNase A + O2 | Trypanosoma brucei | - |
RNase A disulfide + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dithiothreitol + O2 | - |
Trypanosoma brucei | dithiothreitol disulfide + H2O2 | - |
r | |
glutathione + O2 | - |
Trypanosoma brucei | glutathione disulfide + H2O2 | - |
r | |
additional information | redox potentials of TbQSOX-bound FAD and of the CIIIXXCIV proximal disulfide, overview. Determining the redox potential of the CIXXCII motif requires isolating the redox-active TRX domain from the HRR-ERV domains to prevent transfer of reducing equivalents from the CIXXCII dithiol to the CIIIXXCIV and FAD centers of TbQSOX, overview | Trypanosoma brucei | ? | - |
? | |
RNase A + O2 | - |
Trypanosoma brucei | RNase A disulfide + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | domain organization of TbQSOX together with key catalytic steps deduced from studies of both metazoan and protist QSOXs, overview | Trypanosoma brucei |
Synonyms | Comment | Organism |
---|---|---|
QSOX | - |
Trypanosoma brucei |
quiescin sulfhydryl oxidase | - |
Trypanosoma brucei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Trypanosoma brucei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.79 | - |
RNAse A | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
0.9 | - |
dithiothreitol | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
11 | - |
RNAse A | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
13.7 | - |
dithiothreitol | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
21.9 | - |
RNAse A | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei | |
45.1 | - |
dithiothreitol | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Trypanosoma brucei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | reduction of the oxidizing FAD cofactor is followed by the strongly favorable reduction of molecular oxygen, mixed disulfide bond formation is accompanied by the generation of a charge transfer complex with the flavin cofactor. Generation of a 5-deaza-FAD-substituted enzyme by reconstituting the apoprotein with the flavin analogue. Redox potentials of TbQSOX-bound FAD and of the CIIIXXCIV proximal disulfide, overview | Trypanosoma brucei |
General Information | Comment | Organism |
---|---|---|
evolution | mechanistic parallels between the eukaryotic QSOX enzymes and the DsbA/B system catalyzing disulfide bond generation in the bacterial periplasm are detected suggesting that the strategy of linked disulfide exchanges may be exploited in other catalysts of oxidative protein folding | Trypanosoma brucei |
additional information | in the redox center, CXXC motif of the thioredoxin domain is comparatively oxidizing, consistent with an ability to transfer disulfide bonds to a broad range of thiol substrates. In contrast, the proximal CXXC disulfide in the ERV (essential for respiration and vegetative growth) domain of TbQSOX is strongly reducing, representing a major apparent thermodynamic barrier to overall catalysis. Reduction of the oxidizing FAD cofactor is followed by the strongly favorable reduction of molecular oxygen, role of a mixed disulfide intermediate between thioredoxin and ERV domains, overview. Mixed disulfide bond formation is accompanied by the generation of a charge transfer complex with the flavin cofactor providing thermodynamic coupling among the three redox centers of QSOX and avoids the strongly uphill mismatch between the formal potentials of the thioredoxin and ERV disulfides. Domain organization of TbQSOX together with key catalytic steps deduced from studies of both metazoan and protist QSOXs, overview | Trypanosoma brucei |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
19.75 | - |
RNAse A | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
30 | - |
dithiothreitol | mutant CIPHCII, pH 7.0, 25°C | Trypanosoma brucei | |
34.4 | - |
RNAse A | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
60.8 | - |
RNAse A | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei | |
62.3 | - |
dithiothreitol | mutant CIGPCII, pH 7.0, 25°C | Trypanosoma brucei | |
265.3 | - |
dithiothreitol | wild-type enzyme, pH 7.0, 25°C | Trypanosoma brucei |