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Literature summary for 1.1.1.44 extracted from
- The 6-phosphogluconate dehydrogenase from Trypanosoma cruzi: the absence of two inter-subunit salt bridges as a reason for enzyme instability (2004), Mol. Biochem. Parasitol., 133, 197-207.
Application
| Application | Comment | Organism |
|---|---|---|
| pharmacology | the enzyme is a target for inhibitor development for usage as drugs against African Trypanosomiasis | Trypanosoma cruzi |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis, functional overexpression of soluble His-tagged wild-type and mutant enzymes in Escherichia coli | Trypanosoma cruzi |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| V244D/C257R | site-directed mutagenesis, unaltered reaction kinetics and increased stability, due introduction of 2 salt bridges by the double-mutation, compared to the wild-type enzyme | Trypanosoma cruzi |
General Stability
| General Stability | Organism |
|---|---|
| a single step of freezing and thawing reduces the enzyme activity by 90% | Trypanosoma cruzi |
| low stability due to absence of inter-subunit salt bridges as compared to the enzyme from Trypanosoma brucei | Trypanosoma cruzi |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0046 | - |
NADP+ | pH 7.5, 30°C, purified native wild-type enzyme | Trypanosoma cruzi |  |
| 0.0059 | - |
NADP+ | pH 7.5, 30°C, purified recombinant mutant enzyme | Trypanosoma cruzi | |
| 0.022 | - |
6-phosphono-D-gluconate | pH 7.5, 30°C, purified native wild-type enzyme | Trypanosoma cruzi | |
| 0.0222 | - |
6-phosphono-D-gluconate | pH 7.5, 30°C, purified recombinant mutant enzyme | Trypanosoma cruzi | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 51400 | - |
2 * 51980, sequence determination, 2 * 51400, recombinant enzyme, SDS-PAGE | Trypanosoma cruzi |
| 51980 | - |
2 * 51980, sequence determination, 2 * 51400, recombinant enzyme, SDS-PAGE | Trypanosoma cruzi |
| 93000 | - |
recombinant enzyme, gel filtration | Trypanosoma cruzi |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Trypanosoma cruzi | Q6WAT5 | CL Brener clone | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme by partially 27fold ion exchange, affinity and size exclusion chromatography, His-tagged recombinant wild-type and mutant enzymes by nickel affinity chromatography | Trypanosoma cruzi |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| epimastigote | - |
Trypanosoma cruzi | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 32 | - |
purified recombinant enzyme | Trypanosoma cruzi |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 6-phosphono-D-gluconate + NADP+ | - |
Trypanosoma cruzi | D-ribulose 5-phosphate + CO2 + NADPH | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 51980, sequence determination, 2 * 51400, recombinant enzyme, SDS-PAGE | Trypanosoma cruzi |
| More | three-dimensional structure modelling, low stability due to absence of inter-subunit salt bridges as compared to the enzyme from Trypanosoma brucei, the monomeric enzyme is inactive | Trypanosoma cruzi |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 6-phosphonogluconate dehydrogenase | - |
Trypanosoma cruzi |
| 6PGDH | - |
Trypanosoma cruzi |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Trypanosoma cruzi |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 98 | - |
NADP+ | pH 7.5, 30°C, purified native wild-type enzyme | Trypanosoma cruzi | |
| 98 | - |
6-phosphono-D-gluconate | pH 7.5, 30°C, purified native wild-type enzyme | Trypanosoma cruzi | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.4 | - |
- |
Trypanosoma cruzi |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 8.4 | 9.8 | purified native wild-type enzyme, loss of 30% activity within 10 min | Trypanosoma cruzi |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | - |
Trypanosoma cruzi | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Trypanosoma cruzi | electric focusing | - |
6.9 |
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