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Literature summary for 3.6.1.11 extracted from
- NUDIX hydrolases with inorganic polyphosphate exo- and endopolyphosphatase activities in the glycosome, cytosol and nucleus of Trypanosoma brucei (2019), Biosci. Rep., 39, BSR20190894 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus (DE3) | Trypanosoma brucei brucei |
| recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3), recombinant overexpression in Trypanosoma brucei procyclic 29-13 TetR/T7RNAP cell line | Trypanosoma brucei brucei |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | - |
Trypanosoma brucei brucei |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0465 | - |
ADP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 0.0781 | - |
ATP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 0.0822 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 0.1256 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 0.1494 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 0.2005 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytosol | - |
Trypanosoma brucei brucei | 5829 | - |
| glycosome | - |
Trypanosoma brucei brucei | 20015 | - |
| nucleus | - |
Trypanosoma brucei brucei | 5634 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | activates | Trypanosoma brucei brucei | |
| Mg2+ | required | Trypanosoma brucei brucei | |
| Mn2+ | required | Trypanosoma brucei brucei | |
| Mn2+ | activates | Trypanosoma brucei brucei | |
| additional information | enzyme TbNH2 is able to hydrolyze polyphosphate with Mg2+ and Co2+ as cofactors, but has lower activity with Mn2+ | Trypanosoma brucei brucei | |
| additional information | Mg2+ or Mn2+ are the preferred cofactors while no activity is detected with Co2+ | Trypanosoma brucei brucei |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| polyphosphate60 + H2O | Trypanosoma brucei brucei | - |
polyphosphate59 + phosphate | - |
? | |
| polyphosphate60 + H2O | Trypanosoma brucei brucei 927/4 GUTat10.1 | - |
polyphosphate59 + phosphate | - |
? | |
| polyphosphate700 + H2O | Trypanosoma brucei brucei | - |
polyphosphate699 + phosphate | - |
? | |
| polyphosphate700 + H2O | Trypanosoma brucei brucei 927/4 GUTat10.1 | - |
polyphosphate699 + phosphate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Trypanosoma brucei brucei | Q57Z14 | - |
- |
| Trypanosoma brucei brucei | Q583R7 | - |
- |
| Trypanosoma brucei brucei 927/4 GUTat10.1 | Q57Z14 | - |
- |
| Trypanosoma brucei brucei 927/4 GUTat10.1 | Q583R7 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus (DE3) by nickel affinity chromatography and dialysis | Trypanosoma brucei brucei |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ADP + H2O | - |
Trypanosoma brucei brucei | AMP + phosphate | - |
? | |
| ADP + H2O | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | AMP + phosphate | - |
? | |
| ATP + 2 H2O | - |
Trypanosoma brucei brucei | AMP + 2 phosphate | - |
? | |
| ATP + 2 H2O | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | AMP + 2 phosphate | - |
? | |
| additional information | TbNH2 is an exopolyphosphatase with higher activity on short chain polyphosphates. TbNH2 can dephosphorylate ATP and ADP but with lower affinity than for polyphosphate. No activity with 5-diphosphoinositol pentakisphosphate (5-IP7). TbNH2 has a higher affinity for polyphosphate700 than for polyphosphate60. TbNH2 has activity to release the gamma and beta phosphates from ATP and ADP | Trypanosoma brucei brucei | ? | - |
- |
|
| additional information | TbNH4 is an endo- and exopolyphosphatase that has similar activity on polyphosphate of various chain sizes. The enzyme has a higher affinity for polyphosphate60 than for polyphosphate700. No activity with 5-diphosphoinositol pentakisphosphate (5-IP7). Malachite green assay | Trypanosoma brucei brucei | ? | - |
- |
|
| additional information | TbNH2 is an exopolyphosphatase with higher activity on short chain polyphosphates. TbNH2 can dephosphorylate ATP and ADP but with lower affinity than for polyphosphate. No activity with 5-diphosphoinositol pentakisphosphate (5-IP7). TbNH2 has a higher affinity for polyphosphate700 than for polyphosphate60. TbNH2 has activity to release the gamma and beta phosphates from ATP and ADP | Trypanosoma brucei brucei 927/4 GUTat10.1 | ? | - |
- |
|
| additional information | TbNH4 is an endo- and exopolyphosphatase that has similar activity on polyphosphate of various chain sizes. The enzyme has a higher affinity for polyphosphate60 than for polyphosphate700. No activity with 5-diphosphoinositol pentakisphosphate (5-IP7). Malachite green assay | Trypanosoma brucei brucei 927/4 GUTat10.1 | ? | - |
- |
|
| polyphosphate60 + H2O | - |
Trypanosoma brucei brucei | polyphosphate59 + phosphate | - |
? | |
| polyphosphate60 + H2O | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | polyphosphate59 + phosphate | - |
? | |
| polyphosphate700 + H2O | - |
Trypanosoma brucei brucei | polyphosphate699 + phosphate | - |
? | |
| polyphosphate700 + H2O | - |
Trypanosoma brucei brucei 927/4 GUTat10.1 | polyphosphate699 + phosphate | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| exopolyphosphatase | - |
Trypanosoma brucei brucei |
| More | cf. EC 3.6.1.62 and EC 3.6.1.10 | Trypanosoma brucei brucei |
| Nudix hydrolase | - |
Trypanosoma brucei brucei |
| Tb927.5.4350 | - |
Trypanosoma brucei brucei |
| Tb927.6.2670 | - |
Trypanosoma brucei brucei |
| TbDcp2 | - |
Trypanosoma brucei brucei |
| TbNH2 | - |
Trypanosoma brucei brucei |
| TbNH4 | - |
Trypanosoma brucei brucei |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Trypanosoma brucei brucei |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2.512 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 12.555 | - |
ADP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 16.39 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 22.649 | - |
ATP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 27.95 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 44.11 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
assay at | Trypanosoma brucei brucei |
| 8 | - |
- |
Trypanosoma brucei brucei |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the Nudix superfamily (Pfam PF00293) is found in archaea, bacteria, eukaryotes and viruses and includes pyrophosphohydrolases of nucleotide sugars and alcohols, nucleoside and deoxynucleoside triphosphates ([d]NTPs), dinucleoside polyphosphates, dinucleotide coenzymes and capped RNAs. Trypanosoma brucei has five Nudix hydrolases | Trypanosoma brucei brucei |
| physiological function | NHs can participate in polyphosphate homeostasis and therefore may help control polyphosphate levels in glycosomes, cytosol and nuclei of Trypanosoma brucei. Endopolyphosphatase (PPN) activity cleaves internal phosphoanhydride bonds generating shorter polyphosphate molecules. Nudix hydrolase 4 (TbNH4 or TbDcp2) is a mRNA de-capping enzyme that removes the 5' cap from processed mRNAs, EC 3.6.1.62 | Trypanosoma brucei brucei |
| physiological function | NHs can participate in polyphosphate homeostasis and therefore may help control polyphosphate levels in glycosomes, cytosol and nuclei of Trypanosoma brucei. Hydrolysis of polyphosphate with release of phosphate occurs by the activity of a cytosolic exopolyphosphatase (PPX) | Trypanosoma brucei brucei |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 20000 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 110000 | - |
polyphosphate700 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 220000 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 270000 | - |
ATP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 290000 | - |
ADP | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei | |
| 340000 | - |
polyphosphate60 | pH 7.4, 37°C, recombinant enzyme | Trypanosoma brucei brucei |
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