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Literature summary for 3.6.1.53 extracted from

  • Canales, J.; Fernandez, A.; Ribeiro, J.M.; Cabezas, A.; Rodrigues, J.R.; Cameselle, J.C.; Costas, M.J.
    Mn2+-dependent ADP-ribose/CDP-alcohol pyrophosphatase: a novel metallophosphoesterase family preferentially expressed in rodent immune cells (2008), Biochem. J., 413, 103-113.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 cells Rattus norvegicus
expression as GST-fusion protein Rattus norvegicus

Crystallization (Commentary)

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

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 [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

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 [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

Organism UniProt Comment Textmining
Mus musculus Q99KS6
-
-
Rattus norvegicus Q5M886
-
-
Rattus norvegicus Q5M886 bifunctional Mn2+-dependent ADP-ribose/CDP-alcohol diphosphatase
-

Purification (Commentary)

Purification (Comment) Organism
glutathione-Sepharose column chromatography Rattus norvegicus
recombinant GSt-fusion protein, in-column proteolysis of fusion Rattus norvegicus

Source Tissue

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

Storage Stability Organism
-20°C, 1 mM EDTA, stable for months Rattus norvegicus

Substrates and Products (Substrate)

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

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

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 [1/s]

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

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 Range

pH Minimum pH Maximum Comment Organism
6 10
-
Rattus norvegicus

pH Stability

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