Cloned (Comment) | Organism |
---|---|
expression of GST-tagged nezyme in Escherichia coli | Francisella tularensis |
expression of the His-tagged enzyme in Escherichia coli strain Bl21 | Synechocystis sp. |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme in complex with the product AMP and Mn2+ ions in its Nudix active site, sitting drop vapor diffusion method, 20°C, 0.001 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 0.1 M Tris, pH 7.5, 200 mM MgCl2, and 19% PEG 3350, with or without 30 mM AMP, equilibration against the reservoir, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement method | Francisella tularensis |
purified recombinant His-tagged enzyme in complex with co-purified NAD and diphosphate complexed in the NadM-domain active site, and with ADPR substrate complexed in the Nudix-domain, hanging drop vapor diffusion method, 0.0015 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 100 mM Tris, pH 7.5, and 1.5 M Li2SO4, 20°C, 3 days to 2 weeks, X-ray diffraction structure determination and analysis at 2.6 A resolution, selenomethionyl MAD phasing method | Synechocystis sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Francisella tularensis | |
Mg2+ | - |
Synechocystis sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP-ribose + H2O | Francisella tularensis | - |
AMP + D-ribose 5-phosphate | - |
? | |
ADP-ribose + H2O | Synechocystis sp. | - |
AMP + D-ribose 5-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Francisella tularensis | Q5NHR1 | - |
- |
Synechocystis sp. | Q55928 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, the tag is cleaved off by TEV protease, and the detagged protein is further purified by anion exchange chromatography | Francisella tularensis |
recombinant His-tagged enzyme from Escherichia coli strain Bl21 by nickel affinity and anion exchange chromatography, and gel filtration | Synechocystis sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ADP-D-ribose + H2O = AMP + D-ribose 5-phosphate | substrate binding mode in the active site of the ADPRase domain and catalytic mechanism | Francisella tularensis | |
ADP-D-ribose + H2O = AMP + D-ribose 5-phosphate | substrate binding mode in the active site of the ADPRase domain and catalytic mechanism | Synechocystis sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP-ribose + H2O | - |
Francisella tularensis | AMP + D-ribose 5-phosphate | - |
? | |
ADP-ribose + H2O | - |
Synechocystis sp. | AMP + D-ribose 5-phosphate | - |
? | |
ADP-ribose + H2O | substrate binding structure, overview | Francisella tularensis | AMP + D-ribose 5-phosphate | - |
? | |
ADP-ribose + H2O | substrate binding structure, overview | Synechocystis sp. | AMP + D-ribose 5-phosphate | - |
? | |
additional information | the enzyme is a bifunctional NMN adenylyltransferase/ADP-ribose diphosphatase, NMN specificity of the enzyme, overview | Francisella tularensis | ? | - |
? | |
additional information | the enzyme is a bifunctional NMN adenylyltransferase/ADP-ribose diphosphatase, NMN specificity of the enzyme, overview | Synechocystis sp. | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | ADPRase Nudix domain of NadM-Nudix and bacterial monofunctional ADPRase are connected by a long alpha helix with ADPRase active site facing away from the NMNATase domain, while the NMNATase active site opens partially toward the ADPRase domain, structure of the ADPRase domain, overview | Francisella tularensis |
More | ADPRase Nudix domain of NadM-Nudix and bacterial monofunctional ADPRase are connected by a long alpha helix with ADPRase active site facing away from the NMNATase domain, while the NMNATase active site opens partially toward the ADPRase domain, structure of the ADPRase domain, overview | Synechocystis sp. |
Synonyms | Comment | Organism |
---|---|---|
NadM-Nudix | - |
Francisella tularensis |
NadM-Nudix | - |
Synechocystis sp. |
NMN adenylyltransferase/ADP-ribose pyrophosphatase | - |
Francisella tularensis |
NMN adenylyltransferase/ADP-ribose pyrophosphatase | - |
Synechocystis sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Francisella tularensis |
37 | - |
assay at | Synechocystis sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.2 | - |
assay at | Francisella tularensis |
8.2 | - |
assay at | Synechocystis sp. |