Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 cells | Rattus norvegicus |
expression as GST-fusion protein | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
homology modeling based on crystal structure of the Danio rerio protein, Protein Data Base entry 2NXF, and docking of ADP-ribose | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | incubation with 2.5 mM EDTA does not affect ADPRibase-Mn activity; not inhibitory: EDTA up to 2.5 mM | Rattus norvegicus | |
o-phenanthroline | 5 mM causes near full inactivation in 1.5 h | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.039 | - |
ADP-ribose | pH 7.5, 37°C | Rattus norvegicus | |
0.039 | - |
ADP-ribose | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
0.201 | - |
ADP | pH 7.5, 37°C | Rattus norvegicus | |
0.201 | - |
ADP | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
0.3 | - |
CDP-choline | pH 7.5, 37°C | Rattus norvegicus | |
0.3 | - |
CDP-choline | recombinant, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
0.304 | - |
CDP-glycerol | pH 7.5, 37°C | Rattus norvegicus | |
0.304 | - |
CDP-glycerol | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
1.422 | - |
CDP-ethanolamine | pH 7.5, 37°C | Rattus norvegicus | |
1.422 | - |
CDP-ethanolamine | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required. Concentration of MnCl2 giving the half-maximal rate is 0.001-0.004 mM. Mn2+ cannot be replaced by Mg2+ | Rattus norvegicus | |
Mn2+ | dependent on, ADPRibase-Mn is inactive without added Mn2+, and it shows a high affinity for the cation, since saturation is observed at low-micromolar concentrations of Mn2+ well below the substrate concentration (0.5 mM), in no case can Mg2+ replace Mn2+ | Rattus norvegicus | |
additional information | Mg2+ cannot replace Mn2+ | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
gel filtration | Rattus norvegicus |
38000 | - |
SDS-PAGE | Rattus norvegicus |
39360 | - |
calculated from amino acid sequence | Rattus norvegicus |
40000 | - |
1 * 39360, calculated, 1 * 40000, SDS-PAGE, enzyme plus N-terminal extension GPLGSPNS left after proteolysis of recombinant fusion protein | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | Q99KS6 | - |
- |
Rattus norvegicus | Q5M886 | - |
- |
Rattus norvegicus | Q5M886 | bifunctional Mn2+-dependent ADP-ribose/CDP-alcohol diphosphatase | - |
Purification (Comment) | Organism |
---|---|
glutathione-Sepharose column chromatography | Rattus norvegicus |
recombinant GSt-fusion protein, in-column proteolysis of fusion | Rattus norvegicus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Rattus norvegicus | - |
additional information | preferential expression in immune cells | Mus musculus | - |
skeletal muscle | - |
Rattus norvegicus | - |
spleen | ADPRibase-Mn activity in spleen is 2.5-5fold higher than in liver and muscle | Rattus norvegicus | - |
splenocyte | - |
Rattus norvegicus | - |
thymus | ADPRibase-Mn activity in thymus is 2.5-5fold higher than in liver and muscle | Rattus norvegicus | - |
Storage Stability | Organism |
---|---|
-20°C, 1 mM EDTA, stable for months | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + H2O | 27% activity at 0.5 mM substrate compared to ADP-ribose | Rattus norvegicus | ? | - |
? | |
ADP + H2O | 26% of the activity with ADP-ribose | Rattus norvegicus | AMP + phosphate | - |
? | |
ADP-ribose + H2O | - |
Rattus norvegicus | AMP + D-ribose 5-phosphate | - |
? | |
ADP-ribose + H2O | 100% activity at 0.5 mM substrate | Rattus norvegicus | AMP + D-ribose 5-phosphate | - |
? | |
CDP-choline + H2O | 156% of the activity with ADP-ribose | Rattus norvegicus | CMP + phosphocholine | - |
? | |
CDP-choline + H2O | 140% activity at 0.5 mM substrate compared to ADP-ribose | Rattus norvegicus | CMP + phosphocholine | - |
? | |
CDP-ethanolamine + H2O | 84% of the activity with ADP-ribose | Rattus norvegicus | CMP + phosphoethanolamine | - |
? | |
CDP-ethanolamine + H2O | 89% activity at 0.5 mM substrate compared to ADP-ribose | Rattus norvegicus | ? | - |
? | |
CDP-glycerol + H2O | 103% of the activity with ADP-ribose | Rattus norvegicus | CMP + phosphoglycerol | - |
? | |
CDP-glycerol + H2O | 89% activity at 0.5 mM substrate compared to ADP-ribose | Rattus norvegicus | ? | - |
? | |
additional information | ADP-glucose, UDP-glucose, CDP-glucose, CDP, CMP, and AMP are not hydrolysed | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 39360, calculated, 1 * 40000, SDS-PAGE, enzyme plus N-terminal extension GPLGSPNS left after proteolysis of recombinant fusion protein | Rattus norvegicus |
monomer | 1 * 30000, gel filtration | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
ADPRibase-I | isozyme | Rattus norvegicus |
ADPRibase-II | isozyme | Rattus norvegicus |
ADPRibase-Mn | - |
Rattus norvegicus |
Mn2+-dependent ADP-ribose/CDP-alcohol pyrophosphatase | - |
Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.4 | - |
ADP | pH 7.5, 37°C | Rattus norvegicus | |
4.4 | - |
ADP | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
12.8 | - |
ADP-ribose | pH 7.5, 37°C | Rattus norvegicus | |
12.8 | - |
ADP-ribose | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 01 mM MnCl2, at 37°C | Rattus norvegicus | |
28.1 | - |
CDP-glycerol | pH 7.5, 37°C | Rattus norvegicus | |
28.1 | - |
CDP-glycerol | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
28.5 | - |
CDP-choline | pH 7.5, 37°C | Rattus norvegicus | |
28.5 | - |
CDP-choline | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus | |
41.3 | - |
CDP-ethanolamine | pH 7.5, 37°C | Rattus norvegicus | |
41.3 | - |
CDP-ethanolamine | recombinant enzyme, in 50 mM Tris-HCl (pH 7.5), 0.1 mM MnCl2, at 37°C | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | activity towards ADP- and CDP-alcohols | Rattus norvegicus |
8 | 8.5 | - |
Rattus norvegicus |
8 | 8.5 | ADP-ribose hydrolase activity | Rattus norvegicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | - |
Rattus norvegicus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | the activities towards ADP and CDP-alcohols are highest at pH 6-8 and decrease abruptly at higher pH | Rattus norvegicus |