Information on EC 3.1.3.5 - 5'-nucleotidase and Organism(s) Homo sapiens

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Homo sapiens


The taxonomic range for the selected organisms is: Homo sapiens

The enzyme appears in selected viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.1.3.5
-
RECOMMENDED NAME
GeneOntology No.
5'-nucleotidase
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
adenosine nucleotides degradation I
-
-
adenosine nucleotides degradation II
-
-
guanosine nucleotides degradation I
-
-
guanosine nucleotides degradation II
-
-
guanosine nucleotides degradation III
-
-
inosine 5'-phosphate degradation
-
-
NAD salvage pathway III (to nicotinamide riboside)
-
-
pyridine nucleotide cycling (plants)
-
-
tunicamycin biosynthesis
-
-
UTP and CTP dephosphorylation I
-
-
NAD metabolism
-
-
purine metabolism
-
-
Purine metabolism
-
-
Pyrimidine metabolism
-
-
Nicotinate and nicotinamide metabolism
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
SYSTEMATIC NAME
IUBMB Comments
5'-ribonucleotide phosphohydrolase
Wide specificity for 5'-nucleotides.
CAS REGISTRY NUMBER
COMMENTARY hide
9027-73-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
cytosolic 5'-nucleotidase cN-II is implicated in drug resistance in patients and dephosphorylates 5-fluoro-dUMP, 3'-azido-3-dTMP, 2'-chloro-2'-deoxyAMP, and related compounds, which represent the monophosphorylated forms of nucleoside prodrugs
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-chloro-2'-deoxyAMP + H2O
2'-chloro-2'-deoxyadenosine + phosphate
show the reaction diagram
-
cNII
-
-
?
2-chloro-2'-deoxyadenosine monophosphate + H2O
2-chloro-2'-deoxyadenosine + phosphate
show the reaction diagram
39% of the activity with 5'-AMP
-
?
3'-azido-3'deoxy-dTMP + H2O
3'-azido-3'-deoxythymidine + phosphate
show the reaction diagram
-
cNII
-
-
?
5'-AMP + H2O
adenosine + phosphate
show the reaction diagram
5'-CMP + H2O
cytidine + phosphate
show the reaction diagram
5'-dAMP + H2O
deoxyadenosine + phosphate
show the reaction diagram
5'-dCMP + H2O
2'-deoxycytidine + phosphate
show the reaction diagram
5'-dCMP + H2O
deoxycytidine + phosphate
show the reaction diagram
5'-dGMP + H2O
deoxyguanosine + phosphate
show the reaction diagram
5'-dIMP + H2O
deoxyinosine + phosphate
show the reaction diagram
5'-dTMP + H2O
deoxythymidine + phosphate
show the reaction diagram
5'-dUMP + H2O
deoxyuridine + phosphate
show the reaction diagram
5'-dUMP + H2O
deoxyuridne + phosphate
show the reaction diagram
-
i.e. 2'-deoxyuridine 5'-phosphate
-
?
5'-GMP + H2O
guanosine + phosphate
show the reaction diagram
5'-IMP + H2O
deoxyinosine + phosphate
show the reaction diagram
61% of the activity with 5'-AMP
-
?
5'-IMP + H2O
inosine + phosphate
show the reaction diagram
5'-TMP + H2O
thymidine + phosphate
show the reaction diagram
5'-UMP + H2O
uridine + phosphate
show the reaction diagram
5'-XMP + H2O
xanthosine + phosphate
show the reaction diagram
5-fluoro-dUMP + H2O
5-fluoro-uridine + phosphate
show the reaction diagram
-
cNII
-
-
?
9-beta-D-arabinosyl-2-fluoroadenine monophosphate + H2O
9-beta-D-arabinosyl-2-fluoroadenine + phosphate
show the reaction diagram
-
-
-
-
?
dAMP + H2O
deoxyadenosine + phosphate
show the reaction diagram
91% of the activity with 5'-AMP
-
?
dCMP + H2O
deoxycytidine + phosphate
show the reaction diagram
10% of the activity with 5'-AMP
-
?
dGMP + H2O
deoxyguanosine + phosphate
show the reaction diagram
75% of the activity with 5'-AMP
-
?
dIMP + H2O
deoxyinosine + phosphate
show the reaction diagram
84% of the activity with 5'-AMP
-
?
dUMP + H2O
deoxyuridine + phosphate
show the reaction diagram
7% of the activity with 5'-AMP
-
?
FAD + H2O
?
show the reaction diagram
-
microsomal enzyme
-
-
?
IMP + H2O
inosine + phosphate
show the reaction diagram
-
-
-
-
?
inosine + 5'-IMP
5'-IMP + inosine
show the reaction diagram
-
-
-
?
methylthio-IMP + H2O
? + phosphate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
show the reaction diagram
-
-
-
-
?
ribavirin + 5'-IMP
?
show the reaction diagram
better substrate than viramidine
-
-
?
thio-GMP + H2o
6-thioguanosine + phosphate
show the reaction diagram
-
-
-
-
?
thio-IMP + H2O
6-thioinosine + phosphate
show the reaction diagram
-
-
-
-
?
viramidine + 5'-IMP
?
show the reaction diagram
worse substrate than ribavirin
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
5'-AMP + H2O
adenosine + phosphate
show the reaction diagram
5'-CMP + H2O
cytidine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dCMP + H2O
2'-deoxycytidine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dGMP + H2O
deoxyguanosine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dIMP + H2O
deoxyinosine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dTMP + H2O
deoxythymidine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dUMP + H2O
deoxyuridine + phosphate
show the reaction diagram
-
-
-
-
?
5'-GMP + H2O
guanosine + phosphate
show the reaction diagram
-
-
-
-
?
5'-IMP + H2O
inosine + phosphate
show the reaction diagram
-
-
-
-
?
5'-UMP + H2O
uridine + phosphate
show the reaction diagram
-
-
-
-
?
5'-XMP + H2O
xanthosine + phosphate
show the reaction diagram
-
-
-
-
?
IMP + H2O
inosine + phosphate
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
-
-
Zn2+
60% recovery of initial activity with EDTA-treated enzyme
additional information
divalent metal ion is required; no effect: Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-5-(2-(E)-phenylvinyl)-cytosine
-
-
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-5-phenyl-cytosine
-
-
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-cytosine
-
-
1-amino-4-bromo-9,10-dioxo-9,10-dihydroanthracene-2-sulfonic acid
-
-
2,2'-(2-(2-((2S,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-carboxamido)ethylamino)-2-oxoethylazanediyl) diacetic acid
nucleotide-derived inhibitor
2-fluoroadenine-9-beta-D-arabinofuranoside
1 mM, 70% inhibition with 5 mM 5'-AMP as substrate
4-(3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonic acid
-
compound shows high cytotoxicity on H157 cells, with about 72% at 100 mM final concentration
5'-deoxy-5'-S-isobutylthioadenosine
-
-
5-ethynyldideoxyuridine
-
selective inhibitor
6-amino-4-hydroxynaphthalene-2-sulfonic acid
-
most potent inhibitor of the screen
6-N,N-diethyl-D-beta-gamma-dibromomethylene adenosine triphosphate
originally named FPL 67156 or ARL 67156, 28% inhibition at 0.1 mM
6-Thiopurine
-
-
adenosine 5'-(alpha,beta-methylene)diphosphate
adenosine 5'-(alpha,beta-methylene)triphosphate
-
-
alpha,beta-methylene adenosine 5'-diphosphate
-
specific inhibitor
alpha,beta-methylene adenosine-5'-diphosphate
-
-
alpha,beta-methylene-ADP
alpha/beta-methylene-ADP
-
a specific CD73 inhibitor
araCMP
1 mM, 25% inhibition with 5 mM 5'-AMP as substrate
concanavalin A
-
Cu2+
further inhibition of the residual activity in EDTA-treated enzyme
dADP
-
-
dATP
-
-
dCTP
-
-
dGDP
-
-
dGTP
-
-
dITP
-
-
dUTP
-
-
Fe2+
further inhibition of the residual activity in EDTA-treated enzyme
Ly-294002
-
0.01 mM LY-294002 abolishes lipopolysaccharide-induced ecto-5'-nucleotidase activity
NAD+
-
-
NADH
-
-
NADP+
-
-
NADPH
-
-
pentoxifillin
-
-
phosphate
-
-
quercetin
-
-
tenascin C
-
thio-IMP
-
substrate inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-bisphosphoglycerate
2,3-diphosphoglycerate
-
activation
ammonium pyrrolidin dithiocarbamate
-
0.01 mM ammonium pyrrolidin dithiocarbamate increases the basal activity of ecto-5'-nucleotidase by 17% but has no effect on the lipopolysaccharide-induced increase in ecto-5'-nucleotidase activity
D-fructose
-
the activity of 5'-nucleotidase increases with increasing D-fructose concentrations (5-30 mM) in diabetic patients
D-glucose
-
the activity of 5'-nucleotidase increases with increasing D-glucose concentrations (5-30 mM) in diabetic patients
diadenosine 5',5'''-P1,P4-tetraphosphate
-
-
diadenosine polyphosphate
-
allosteric activation
-
Diadenosine tetraphosphate
-
activates cytosolic 5'-nucleotidase II by increase binding and dimer formation, interface binding structure modeling
indomethacin
-
0.1 mM, causes 62% increase of activity in vivo after 48 h
lipopolysaccharide
-
ecto-5'-nucleotidase activity increases after incubation with 10 ng/ml lipopolysaccharide for 24 h but is not changed after shorter incubation times
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0038 - 2.96
5'-AMP
0.002 - 0.0564
5'-CMP
0.035 - 0.114
5'-dAMP
0.023
5'-dCMP
-
-
0.044
5'-dGMP
-
-
0.057
5'-dIMP
-
-
0.017
5'-dUMP
-
-
0.0065 - 0.017
5'-GMP
0.0071 - 12.11
5'-IMP
0.008
5'-TMP
37C, pH not specified in the publication
0.014 - 0.0316
5'-UMP
0.065
5'-XMP
-
-
10
9-beta-D-arabinosyl-2-fluoroadenine monophosphate
-
pH 8.3, 25C
0.0091
AMP
-
at pH 8.5
0.012 - 0.193
dCMP
1.8 - 2.17
Inosine
0.024 - 0.095
ribavirin
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
187
5'-AMP
37C, pH not specified in the publication
202
5'-CMP
37C, pH not specified in the publication
174
5'-dAMP
37C, pH not specified in the publication
132
5'-GMP
37C, pH not specified in the publication
104
5'-IMP
37C, pH not specified in the publication
40
5'-TMP
37C, pH not specified in the publication
289
5'-UMP
37C, pH not specified in the publication
19 - 22
Inosine
0.023 - 0.15
ribavirin
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
48800
5'-AMP
37C, pH not specified in the publication
3600
5'-CMP
37C, pH not specified in the publication
1500
5'-dAMP
37C, pH not specified in the publication
20300
5'-GMP
37C, pH not specified in the publication
14700
5'-IMP
37C, pH not specified in the publication
5000
5'-TMP
37C, pH not specified in the publication
9100
5'-UMP
37C, pH not specified in the publication
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.14
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-5-(2-(E)-phenylvinyl)-cytosine
-
pH 6.5, 37C
2.01
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-5-phenyl-cytosine
-
pH 6.5, 37C
1.74
1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-cytosine
-
pH 6.5, 37C
0.00051
alpha,beta-methylene-ADP
-
in 10 mM HEPES (pH 7.4), 2 mM MgCl2, and 1 mM CaCl2
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.004
1-amino-4-bromo-9,10-dioxo-9,10-dihydroanthracene-2-sulfonic acid
Homo sapiens;
-
pH 7.4, 37C
0.0061
4-(3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl)benzenesulfonic acid
Homo sapiens;
-
pH 7.4, 37C
0.02
5-ethynyldideoxyuridine
Homo sapiens;
-
-
0.00132
6-amino-4-hydroxynaphthalene-2-sulfonic acid
Homo sapiens;
-
pH 7.4, 37C
0.0453
quercetin
Homo sapiens;
-
pH 7.4, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17.1
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
-
microsomal enzyme
6.5
-
cytoplasmic enzyme, substrate 5'-IMP
7
-
substrate thio-IMP
7.5
-
substrates IMP, methylthio-IMP
8
-
substrate thio-GMP
8.5
-
substrates GMP, AMP
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
bronchial epithelium, mucosal and serosal epithelial surface
Manually annotated by BRENDA team
-
patients with head and neck cancer
Manually annotated by BRENDA team
-
5'-nucleotidase activity is decreased in seminal plasma from varicocele patients
Manually annotated by BRENDA team
-
5'-nucleotidase activity is decreased in spermatozoa from varicocele patients
Manually annotated by BRENDA team
-
CD73
Manually annotated by BRENDA team
-
CD73 is a lymphocyte surface antigen
Manually annotated by BRENDA team
-
CD73
Manually annotated by BRENDA team
-
the enzyme contributes to function of endothelial cells by inhibiting platelet aggregation in cooperation with ATP diphosphohydrolase, which degrades ADP to AMP
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
cell surface
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
UNIPROT
ORGANISM
Homo sapiens;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47700
x * 47700, calculation from nucleotide sequence
53000
-
4 * 53000, cytoplasmic enzyme, SDS-PAGE
56000
-
x * 56000, membrane-bound enzyme, deglycosylated, SDS-PAGE
57000
-
SDS-PAGE
57300
-
4 * 57300, calculated from amino acid sequence
58000
-
x * 67000, glycosylated enzyme form, x * 58000, deglycosylated enzyme form, SDS-PAGE
65000
x * 65000, truncated form of enzyme
67000
-
x * 67000, glycosylated enzyme form, x * 58000, deglycosylated enzyme form, SDS-PAGE
70000
-
SDS-PAGE
71000
-
homodimeric CD73
73000
-
x * 73000, membrane-bound enzyme, SDS-PAGE under reducing conditions
74000
-
x * 74000, microsomal enzyme, SDS-PAGE
150000
-
-
195000
-
gel filtration
210000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
-
homodimer
-
CD73
homotetramer
-
4 * 57300, calculated from amino acid sequence
tetramer
additional information
-
cN-II is a relatively large enzyme of the structural HAD superfamily, which contains, besides the alpha/beta-Rossmann fold and the four-helix bundle, non-conserved regions involved in effector binding and subunit-subunit interactions
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of crystal structures of the inactive D52N mutant enzyme in apoform or in complexes with 1. 2,3-bisphosphoglycerate, 2. IMP/ATP, 3. IMP/2,3-bisphosphoglycerate, 4. GMP/diadenosine tetraphosphate, 5. dGMP/dATP, or 6. UMP/ATP, overview. Crystallization by sitting drop vapor diffusion method with sitting drops of 300 nl at a protein solution/reservoir solution ratio of 2:1, the reservoir solution contains 0.1 M bicine, pH 9.0, and 10% w/v PEG 6000, with 20 mM MgCl2, 10 mM dATP, and 10 mM dGMP added to the protein solution, 7.7 mg/ml protein, 1-2 weeks at 4C, to obtain the enzyme complexes the crystals are soaked in 0.01 ml drops of reservoir solution supplemented with 10 mM of the corresponding substrate/activator and 20 mM MgCl2 for 90 min, X-ray diffraction structure determination and analysis at A resolution
-
in complex with adenosine, inhibitors 2,2'-(2-(2-((2S,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-carboxamido)ethylamino)-2-oxoethylazanediyl) diacetic acid, baicalin and ADP analog AMPCP. The dimeric enzyme undergoes an extensive 114 conformational switch between the open and closed forms. The dimerization interface is formed by the C-terminal domains and exhibits interchain motions of up to 13. Structural control of the domain movement determines the selectivity for monophosphate nucleotides
molecular modeling study using inhibitor 1-(6-deoxy-6-phosphono-beta-D-allofuranosyl)-5-(2-(E)-phenylvinyl)-cytosine
-
sitting drop vapour diffusion method with 1.8 M of MgSO4 and 0.1 M of Tris, pH 8.5
-
soluble form of human soluble CD73 at 2.2 A resolution, a truncated form of CD73, residues 27549, that retains ecto-5'-nucleotidase activity. Structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56
-
30 min, 44% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni2+-charged His-Trap HP column chromatography and HiLoad 16/60 Superdex 200 gel filtration
-
recombinant enzyme
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli Rosetta2(DE) cells
-
expression in Sf9 insect cells
expression of mutant D52N in Escherichia coli strain BL21(DE)-R3-pRARE2
-
gene NT5E at 6q14-q21 encodes CD73
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
CD73 is highly induced by ischemia and hypoxia, and increased expression in cardiac ischemia is mediated by a positive feedback loop involving adenosine. Though CD73 is mainly regulated transcriptionally, cross linking of CD38, a cell surface glycoprotein with cyclic ADP-ribose hydrolase activity, causes upregulation of CD73 via export of protein to the cell surface that is independent of new transcription
-
markedly increased in the ventricular myocardium in patients with chronic heart failure
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D52N
-
site-directed mutagenesis, catalytically inactive active site residue
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine