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Literature summary extracted from

  • Zebisch, M.; Krauss, M.; Schaefer, P.; Straeter, N.
    Crystallographic evidence for a domain motion in rat nucleoside triphosphate diphosphohydrolase (NTPDase) 1 (2012), J. Mol. Biol., 415, 288-306.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.1.5 expressed in Escherichia coli Rattus norvegicus
3.6.1.5 recombinant expression of the mutant NTPDase1 ECD DELTAMIL in Escherichia coli in soluble form and refolding to the active state Rattus norvegicus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.6.1.5 NTPDase1 that lacks the two transmembrane helices and an additional putative membrane interaction loop, hanging drop vapor diffusion method, using 0.1 M sodium acetate (pH 4.5), 3.6 M sodium chloride Rattus norvegicus
3.6.1.5 purified recombinant wild-type enzyme, free or in complex with decavanadate or heptamolybdate, X-ray diffraction structure determination and analysis at 1.7-2.5 A resolution, modeling Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
3.6.1.5 K257M site-directed mutagenesis Rattus norvegicus
3.6.1.5 additional information replacement of the extracellular domain of Rattus norvegicus NTPDase1, i.e. amino acid sequence 190TQEQSWLNFISDSQKQA206, with the shorter hydrophilic loop found in Homo sapiens NTPDase6, 240KTPGGS245. This NTPDase1 ECD DELTAMIL mutant variant is a soluble NTPDase1 that lacks a putative membrane interaction loop identified between the two lobes of the catalytic domain Rattus norvegicus
3.6.1.5 Y409F site-directed mutagenesis Rattus norvegicus
3.6.1.5 Y413F site-directed mutagenesis Rattus norvegicus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.6.1.5 ammonium heptamolybdate AHM, (NH4)6[Mo7O24] Rattus norvegicus
3.6.1.5 AMP
-
Rattus norvegicus
3.6.1.5 additional information inhibition mechanism of polyoxometallates, overview Rattus norvegicus
3.6.1.5 polytungstate salt POM-1 Na6[H2W12O40] Rattus norvegicus
3.6.1.5 Sodium vanadate Na3VO4 Rattus norvegicus
3.6.1.5 UMP
-
Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6.1.5 additional information
-
additional information kinetics, overview Rattus norvegicus
3.6.1.5 0.0025
-
ADP recombinant mutant EDC K257M, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0025
-
ADP recombinant mutant EDC Y314F, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0047
-
ADP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0051
-
ADP recombinant mutant EDC Y409F, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0059
-
ADP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.009
-
GTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.01
-
UTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0105
-
IDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0108
-
ITP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0113
-
UDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0114
-
GDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0129
-
UTP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0131
-
GDP in presence of 0.1 mM UMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0175
-
UDP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0488
-
GDP in presence of 0.1 mM AMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.149
-
GDP in presence of 0.1 mM AHM, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.1712
-
AMPCPP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.7495
-
TDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.6.1.5 extracellular
-
Rattus norvegicus
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.6.1.5 Cl- the active-site clefts of NTPDase1 contain three chloride ions, one of which is bound to the second phosphate-binding loop, apyrase conserved region 4, ACR4 Rattus norvegicus
3.6.1.5 additional information complex structures with decavanadate and heptamolybdate show that both polyoxometallates bind electrostatically to a loop that is involved in binding of the nucleobase Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.6.1.5 ATP + 2 H2O Rattus norvegicus
-
AMP + 2 phosphate
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.6.1.5 Rattus norvegicus P97687
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.6.1.5 vascular system CD39 and NTPDase1 are the dominant NTPDases Rattus norvegicus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.6.1.5 adenosine 5'-[(alpha,beta)-methyleno] triphosphate + H2O
-
Rattus norvegicus ?
-
?
3.6.1.5 ADP + H2O
-
Rattus norvegicus AMP + phosphate
-
?
3.6.1.5 AMPCPP + H2O low activity Rattus norvegicus ?
-
?
3.6.1.5 ATP + 2 H2O
-
Rattus norvegicus AMP + 2 phosphate
-
?
3.6.1.5 ATP + H2O
-
Rattus norvegicus ADP + phosphate
-
?
3.6.1.5 GDP + H2O
-
Rattus norvegicus GMP + phosphate
-
?
3.6.1.5 GTP + 2 H2O
-
Rattus norvegicus GMP + 2 phosphate
-
?
3.6.1.5 IDP + H2O
-
Rattus norvegicus IMP + phosphate
-
?
3.6.1.5 ITP + 2 H2O
-
Rattus norvegicus IMP + 2 phosphate
-
?
3.6.1.5 ITP + H2O
-
Rattus norvegicus IDP + phosphate
-
?
3.6.1.5 additional information establishment of a kinetic isothermal titration calorimetry assay. Substrate recognition by NTPDase1, overview Rattus norvegicus ?
-
?
3.6.1.5 additional information no hydrolysis of AMPCP Rattus norvegicus ?
-
?
3.6.1.5 TDP + H2O
-
Rattus norvegicus TMP + phosphate
-
?
3.6.1.5 TTP + 2 H2O weak substrate Rattus norvegicus TMP + 2 phosphate
-
?
3.6.1.5 UDP + H2O
-
Rattus norvegicus UMP + phosphate
-
?
3.6.1.5 UTP + 2 H2O
-
Rattus norvegicus UMP + 2 phosphate
-
?
3.6.1.5 UTP + H2O
-
Rattus norvegicus UDP + phosphate
-
?

Synonyms

EC Number Synonyms Comment Organism
3.6.1.5 CD39
-
Rattus norvegicus
3.6.1.5 NTPDase 1
-
Rattus norvegicus
3.6.1.5 NTPDase1
-
Rattus norvegicus
3.6.1.5 nucleoside triphosphate diphosphohydrolase 1
-
Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.6.1.5 25
-
assay at Rattus norvegicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.6.1.5 31.1
-
ADP recombinant mutant EDC Y409F, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 40.5
-
AMPCPP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 50.4
-
ADP recombinant mutant EDC K257M, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 77.3
-
ADP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 79.9
-
IDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 80.2
-
TDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 80.9
-
UDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 80.9
-
GTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 82.6
-
GDP in presence of 0.1 mM AMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 83.2
-
GDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 83.2
-
GDP in presence of 0.1 mM UMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 86.1
-
ADP recombinant mutant EDC Y314F, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 86.9
-
ADP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 93
-
UDP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 103.4
-
UTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 103.7
-
GDP in presence of 0.1 mM AHM, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 105.3
-
ITP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 121.2
-
UTP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.6.1.5 7.4
-
assay at Rattus norvegicus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.6.1.5 0.00859
-
ammonium heptamolybdate recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0268
-
AMP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.0268
-
AMP at pH 7.4 and 25°C Rattus norvegicus
3.6.1.5 0.3345
-
UMP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 0.3345
-
UMP at pH 7.4 and 25°C Rattus norvegicus

General Information

EC Number General Information Comment Organism
3.6.1.5 evolution the enzyme is a member of the eukaryotic NTPDase family Rattus norvegicus
3.6.1.5 additional information influence of transmembrane helix dynamics on activity is achieved by coupling to a domain motion. Active site structure of NTPDase1, overview, closure movement in NTPDases Rattus norvegicus
3.6.1.5 physiological function nucleoside triphosphate diphosphohydrolases are a physiologically important class of membrane-bound ectonucleotidases responsible for the regulation of extracellular levels of nucleotides Rattus norvegicus
3.6.1.5 physiological function the enzyme blocks platelet aggregation and supports blood flow Rattus norvegicus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.6.1.5 110
-
TDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 230
-
AMPCPP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 700
-
GDP in presence of 0.1 mM AHM, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 1700
-
GDP in presence of 0.1 mM AMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 5300
-
UDP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 6100
-
ADP recombinant mutant EDC Y409F, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 6400
-
GDP in presence of 0.1 mM UMP, recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 7200
-
UDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 7300
-
GDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 7600
-
IDP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 9000
-
GTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 9400
-
UTP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 9800
-
ITP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 10300
-
UTP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 14700
-
ADP recombinant mutant EDC DELTA MIL, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 16400
-
ADP recombinant wild-type enzyme, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 20200
-
ADP recombinant mutant EDC K257M, pH 7.4, 25°C Rattus norvegicus
3.6.1.5 26900
-
ADP recombinant mutant EDC Y314F, pH 7.4, 25°C Rattus norvegicus