Information on EC 2.4.2.2 - pyrimidine-nucleoside phosphorylase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
2.4.2.2
-
RECOMMENDED NAME
GeneOntology No.
pyrimidine-nucleoside phosphorylase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2'-deoxyuridine + phosphate = uracil + 2-deoxy-alpha-D-ribose 1-phosphate
show the reaction diagram
molecular basis of catalytic chamber-assisted unfolding and cleavage of human insulin by human insulin-degrading enzyme, IDE utilizes the interaction of its exosite with the N-terminus of the insulin A chain, overview. The unique size, shape, charge distribution, and exosite of the IDE catalytic chamber contribute to its high affinity for insulin
-
-
-
thymidine + phosphate = thymine + 2-deoxy-alpha-D-ribose 1-phosphate
show the reaction diagram
(2)
-
-
-
uridine + phosphate = uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
; (1)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pentosyl group transfer
-
-
-
-
pentosyl group transfer
-
-
pentosyl group transfer
Q5SHF9, -
-
pentosyl group transfer
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
pyrimidine deoxyribonucleosides degradation
-
Pyrimidine metabolism
-
pyrimidine ribonucleosides degradation
-
SYSTEMATIC NAME
IUBMB Comments
pyrimidine-nucleoside:phosphate alpha-D-ribosyltransferase
Unlike EC 2.4.2.3, uridine phosphorylase, and EC 2.4.2.4, thymidine phosphorylase, this enzyme can accept both the ribonucleoside uridine and the 2'-deoxyribonucleosides 2'-deoxyuridine and thymidine [3]. The reaction is reversible, and the enzyme does not distinguish between alpha-D-ribose 1-phosphate and 2-deoxy-alpha-D-ribose 1-phosphate in the synthetic direction.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phosphorylase, pyrimidine nucleoside
-
-
-
-
Py-NPase
-
-
-
-
PYNP
-
-
-
-
pynpase
-
-
-
-
pynpase
-
-
pyrimidine nucleoside phosphorylase
-
-
-
-
pyrimidine nucleoside phosphorylase
-
-
pyrimidine nucleoside phosphorylase
-
-
pyrimidine nucleoside phosphorylase
Q5SHF9
-
pyrimidine ribonucleoside phosphorylase
-
-
-
-
TTHA1771
Q5SHF9
-
CAS REGISTRY NUMBER
COMMENTARY
9055-35-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Enterobacter aerogenes CMCC 45103
-
-
-
Manually annotated by BRENDA team
Geobacillus stearothermophilus JTS 859
JTS 859
-
-
Manually annotated by BRENDA team
Haemophilus influenzae TH 6-2
TH 6-2
-
-
Manually annotated by BRENDA team
strain HB8
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)uracil + phosphate
uracil + 2'-deoxy-2'-fluoro-beta-D-arabinofuranose 1-phosphate
show the reaction diagram
Q72HS4
-
1.6% substrate conversion after 30 min
-
?
1,2,4-triazole-3-carboxamide + alpha-D-ribose 1-phosphate
ribavirin + phosphate
show the reaction diagram
-
-
-
-
-
2'-deoxy-2'-fluorouridine + phosphate
?
show the reaction diagram
-
-
poor activity, 0.4% substrate conversion after 30 min
-
?
2'-deoxy-2'-fluorouridine + phosphate
uracil + 2'-deoxy-2'-fluoro-ribose 1-phosphate
show the reaction diagram
Q72HS4
-
15.6% substrate conversion after 30 min
-
?
4-thiothymidine + phosphate
4-thiothymin + alpha-D-ribose 1-phosphate
show the reaction diagram
-
uridine phosphorylase activity, see also EC 2.4.2.3
-
-
r
4-thiouridine + phosphate
4-thiouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
thymidine phosphorylase activity, see also EC 2.4.2.4
-
-
r
5'-deoxy-5-fluorouridine + phosphate
5-fluorouracil + alpha-D-deoxyribose 1-phosphate
show the reaction diagram
-
-
5-fluorouracil is a chemotherapeutic agent and inactivated by dihydropyrimidine dihydrogenase activity
-
?
5-bromodeoxyuridine + phosphate
5-bromouracil + deoxyribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-bromodeoxyuridine + phosphate
5-bromouracil + deoxyribose 1-phosphate
show the reaction diagram
-
95% of the activity with uridine
-
-
?
5-bromodeoxyuridine + phosphate
5-bromouracil + deoxyribose 1-phosphate
show the reaction diagram
-
74% of the activity with uridine
-
-
?
5-bromodeoxyuridine + phosphate
5-bromouracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-bromouridine + phosphate
5-bromouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
-
5-bromouridine + phosphate
5-bromouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
40% of the activity with uridine
-
-
?
5-bromouridine + phosphate
5-bromouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
95% of the activity with uridine
-
-
?
5-chlorodeoxyuridine + phosphate
5-chlorouracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-fluorodeoxyuridine + phosphate
5-fluorouracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-fluorouridine + phosphate
5-fluorouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-iododeoxyuridine + phosphate
5-iodouracil + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-iododeoxyuridine + phosphate
5-iodouracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-iodouridine + phosphate
5-iodouracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-methyluridine + phosphate
5-methyluracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-methyluridine + phosphate
5-methyluracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
5-methyluridine + phosphate
5-methyluracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
27% of the activity with uridine
-
-
?
5-methyluridine + phosphate
5-methyluracil + alpha-D-ribose 1-phosphate
show the reaction diagram
Geobacillus stearothermophilus JTS 859
-
-
-
-
?
5-trifluoromethyl-2'-deoxyuridine + phosphate
5-trifluoromethyluracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
cytidine + phosphate
cytosine + alpha-D-ribose 1-phosphate
show the reaction diagram
-
no activity
-
-
-
cytidine + phosphate
cytosine + alpha-D-ribose 1-phosphate
show the reaction diagram
-
8% of the activity with uridine
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
12% of the activity with uridine
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
2'-deoxyuridine
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
179% of the activity with uridine, by measuring the conversion of uracil to uridine or deoxyuridine a slight preference for ribose 1-phosphate over deoxyribose 1-phosphate is observed
-
-
r
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
147% of the activity with uridine
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
Geobacillus stearothermophilus JTS 859
-
2'-deoxyuridine
-
-
?
deoxyuridine + phosphate
uracil + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
Haemophilus influenzae TH 6-2
-
147% of the activity with uridine
-
-
?
deoxyuridine + phosphate
uracil + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
pyrimidine nucleoside + phosphate
pyrimidine base + alpha-D-ribose 1-phosphate
show the reaction diagram
-
most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine
or alpha-D-deoxyribose 1-phosphate
-
r
pyrimidine nucleoside + phosphate
pyrimidine base + alpha-D-ribose 1-phosphate
show the reaction diagram
Q5SHF9, -
most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine
or alpha-D-deoxyribose 1-phosphate
-
r
ribofuranosyl thymine + phosphate
thymine + alpha-D-ribose 1-phosphate
show the reaction diagram
Haemophilus influenzae, Haemophilus influenzae TH 6-2
-
-
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
21% of the activity with uridine
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
139% of the activity with uridine
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
99% of the activity with uridine
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
Geobacillus stearothermophilus JTS 859
-
-
-
-
?
thymidine + phosphate
thymine + 2-deoxy-D-ribose 1-phosphate
show the reaction diagram
Haemophilus influenzae TH 6-2
-
99% of the activity with uridine
-
-
?
thymidine + phosphate
thymine + alpha-D-deoxyribose 1-phosphate
show the reaction diagram
Q5SHF9, -
-
-
-
r
thymidine + phosphate
thymine + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
thymidine + phosphate
thymine + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
thymidine + phosphate
thymine + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
Q72HS4
-
-
-
?
thymidine + phosphate
thymine + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
Q5XA29
-
-
-
?
uracil arabinoside + phosphate
uracil + alpha-D-arabinose 1-phosphate
show the reaction diagram
-
-
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
-
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
-
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
preferred substrate
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
Q5XA29
preferred substrate
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
by measuring the conversion of uracil to uridine or deoxyuridine a slight preference for ribose 1-phosphate over deoxyribose 1-phosphate is observed
-
r
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
prefered substrate
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
-
prefered substrate
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
Q72HS4
prefered substrate
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
Geobacillus stearothermophilus JTS 859
-
-
-
-
?
uridine + phosphate
uracil + alpha-D-ribose 1-phosphate
show the reaction diagram
Haemophilus influenzae TH 6-2
-
-
-
-
?
deoxyuridine + phosphate
uracil + 2'-deoxy-D-ribose 1-phosphate
show the reaction diagram
Q5XA29
-
-
-
?
additional information
?
-
-
no activity with deoxycytidine
-
-
-
additional information
?
-
-
reversible phosphorolysis of pyrimidines in the nucleotide synthesis salvage pathway
-
-
-
additional information
?
-
-
predictive profiling of 5-fluorouracil resistance in human cancer cells, overview
-
-
-
additional information
?
-
-, P39142
the function of His82, Arg169, and Lys188 is to stabilize the uridine in a high-energy conformation by means of electrostatic interactions. These residues are involved in catalysis. The function of Asp162 is likely to activate Lys188 for phosphorolytic catalysis through polarization effects
-
-
-
additional information
?
-
-
no substrate: cytidine, deoxycytidine, 2',2'-difluoro-2'-deoxycytidine, 2',2'-difluoro-2'-deoxyuridine, uracil arabinoside, thymine arabinoside, (E)-5-(2-bromovinyl)-deoxyuridine, azidothymidine, stavudine, 6-azauridine
-
-
-
additional information
?
-
Q5XA29
no substrate: cytidine, deoxycytidine, adenosine, inosine, or guanosine
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
pyrimidine nucleoside + phosphate
pyrimidine base + alpha-D-ribose 1-phosphate
show the reaction diagram
-
most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine
or alpha-D-deoxyribose 1-phosphate
-
r
pyrimidine nucleoside + phosphate
pyrimidine base + alpha-D-ribose 1-phosphate
show the reaction diagram
Q5SHF9, -
most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine
or alpha-D-deoxyribose 1-phosphate
-
r
additional information
?
-
-
reversible phosphorolysis of pyrimidines in the nucleotide synthesis salvage pathway
-
-
-
additional information
?
-
-
predictive profiling of 5-fluorouracil resistance in human cancer cells, overview
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3,5-bis-(4-chlorobenzoyl)-alpha-D-ribose 1-phosphate
-
competitive inhibition with regard to inosine
-
5-benzylacyclouridine
-
-
5-chloro-6-(1-[2-iminopyrrolidinyl]methyl)uracil
-
-
-
uridine
-
above 15 mM, competitive inhibition of cleavage of 5-bromodeoxyuridine
7-deazaxanthine
-
-
additional information
-
inhibitory effects between pyrimidines and analogues
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.58
-
2'-Deoxyuridine
-
-
0.25
-
4-thiothymidine
-
pH 7.5, 25C, thymidine phosphorylase activity
0.13
-
4-thiouridine
-
pH 7.5, 25C, uridine phosphorylase activity
0.1
-
5-bromodeoxyuridine
-
-
0.03
-
5-Bromouridine
-
-
0.169
-
5-fluorodeoxyuridine
-
pH 7.6, 37C
0.047
-
5-fluorouridine
-
pH 7.6, 37C
0.144
-
5-iododeoxyuridine
-
pH 7.6, 37C
0.069
-
5-iodouridine
-
pH 7.6, 37C
0.07
-
5-methyluridine
-
-
0.32
-
5-methyluridine
-
-
0.13
-
Deoxyuridine
-
-
0.38
-
phosphate
-
reaction with uridine
0.388
-
phosphate
-
cosubstrate uridine, pH 7.6, 37C
0.797
-
phosphate
-
cosubstrate deoxythymidine, pH 7.6, 37C
0.43
-
thymidine
Q72HS4
pH 7.0, 60C
0.473
-
thymidine
-
pH 7.6, 37C
1.093
-
thymidine
-
pH 7.6, 37C
1.3
-
thymidine
-
pH 7.0, 60C
1.9
-
thymidine
-
-
0.092
-
uridine
-
pH 7.6, 37C
0.15
-
uridine
Q72HS4
pH 7.0, 60C
0.158
-
uridine
Q5XA29
wild-type, pH 7.5, temperature not specified in the publication
0.171
-
uridine
Q5XA29
mutant H169A, pH 7.5, temperature not specified in the publication
0.31
-
uridine
-
pH 7.6, 37C
2.3
-
uridine
-
pH 7.0, 60C
0.578
-
Deoxyuridine
-
pH 7.6, 37C
additional information
-
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
63
-
4-thiothymidine
-
pH 7.5, 25C, thymidine phosphorylase activity
49
-
4-thiouridine
-
pH 7.5, 25C, uridine phosphorylase activity
11.9
-
5-fluorodeoxyuridine
-
pH 7.6, 37C
6.1
-
5-fluorouridine
-
pH 7.6, 37C
9.5
-
5-iododeoxyuridine
-
pH 7.6, 37C
6.4
-
5-iodouridine
-
pH 7.6, 37C
24.6
-
Deoxyuridine
-
pH 7.6, 37C
8.6
-
phosphate
-
cosubstrate uridine, pH 7.6, 37C
17.5
-
phosphate
-
cosubstrate deoxythymidine, pH 7.6, 37C
21.6
-
thymidine
-
pH 7.6, 37C
83
-
thymidine
-
pH 7.0, 60C
679
-
thymidine
Q72HS4
pH 7.0, 60C
1.7
-
uridine
Q5XA29
mutant H169A, pH 7.5, temperature not specified in the publication
8.5
-
uridine
-
pH 7.6, 37C
15
-
uridine
Q5XA29
wild-type, pH 7.5, temperature not specified in the publication
277
-
uridine
-
pH 7.0, 60C
277
-
uridine
Q72HS4
pH 7.0, 60C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
70
-
5-fluorodeoxyuridine
-
pH 7.6, 37C
5235
130
-
5-fluorouridine
-
pH 7.6, 37C
5244
66
-
5-iododeoxyuridine
-
pH 7.6, 37C
5318
93
-
5-iodouridine
-
pH 7.6, 37C
83370
43
-
Deoxyuridine
-
pH 7.6, 37C
9639
22
-
phosphate
-
cosubstrate deoxythymidine, pH 7.6, 37C; cosubstrate uridine, pH 7.6, 37C
27500
46
-
thymidine
-
pH 7.6, 37C
17198
92
-
uridine
-
pH 7.6, 37C
17761
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
additional information
-
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.113
-
3,5-bis-(4-chlorobenzoyl)-alpha-D-ribose 1-phosphate
-
substrate deoxythymidine, pH 7.6, 37C
-
0.128
-
3,5-bis-(4-chlorobenzoyl)-alpha-D-ribose 1-phosphate
-
substrate 5-fluorodeoxyuridine, pH 7.6, 37C
-
0.159
-
3,5-bis-(4-chlorobenzoyl)-alpha-D-ribose 1-phosphate
-
substrate 5-iododeoxyuridine pH 7.6, 37C
-
0.188
-
3,5-bis-(4-chlorobenzoyl)-alpha-D-ribose 1-phosphate
-
substrate uridine, pH 7.6, 37C
-
0.000003
-
5-benzylacyclouridine
-
substrate uridine, pH 7.6, 37C
0.000005
-
5-benzylacyclouridine
-
substrate deoxythymidine, pH 7.6, 37C
0.5
-
5-chloro-6-(1-[2-iminopyrrolidinyl]methyl)uracil
-
substrate deoxythymidine, pH 7.6, 37C; substrate uridine, pH 7.6, 37C
-
0.03
-
7-deazaxanthine
-
substrate deoxythymidine, pH 7.6, 37C
0.06
-
7-deazaxanthine
-
substrate uridine, pH 7.6, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.9
-
Q5XA29
substrate thymidine, pH 7.5, temperature not specified in the publication
8.1
-
Q5XA29
substrate uridine, pH 7.5, temperature not specified in the publication
11.2
-
Q5XA29
substrate deoxyuridine, pH 7.5, temperature not specified in the publication
45
-
-
purified recombinant enzyme
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.9
-
-
deoxynucleoside substrates
7.2
-
-
uridine, thymidine
7.4
-
-
ribonucleosides and uracil arabinoside
7.5
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
12
-
pH 6.0: about 75% of maximal activity, pH 12.0: about 80% of maximal activity
6.9
7.4
-
pH 6.9: optimum, pH 7.4: 75% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
PyNPase level quantified by sandwich ELISA using monoclonal anti-human thimidine phosphorylase antibodies 104B and 232-2, no association between PyNPase level and recurrence, disease-free survival at year 2.2 better with low PyNPase levels and high PyNPase-to-dihydropyrimidine dihydrogenase ratio
Manually annotated by BRENDA team
-
PyNPase level quantified by sandwich ELISA using monoclonal anti-human thimidine phosphorylase antibodies 104B and 232-2
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Staphylococcus aureus (strain COL)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
45400
-
Q5SHF9, -
estimated according to amino acid composition
78000
-
-
sedimentation velocity analysis
85000
-
-
gel filtration
86300
-
Q5SHF9, -
in the dimeric state of the purified protein
92000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 46000, SDS-PAGE
dimer
-
2 * 54000, SDS-PAGE
dimer
Q5SHF9, -
revealed by dynamic light-scattering (DynaPro MS/X) of the purified protein, 20 mg protein/ml, 18C, 20 mM Tris-HCl pH 7.6, 200 mM sodium chloride, particle size distribution with a polydispersity of 18.7%
dimer
Geobacillus stearothermophilus JTS 859
-
2 * 54000, SDS-PAGE
-
dimer
Haemophilus influenzae TH 6-2
-
2 * 46000, SDS-PAGE
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of the enzyme with the substrate analog, pseudouridine, in its active site is solved to 2.1 A
-
hanging-drop vapor diffusion method, crystals of the protein-inhibitor complex with the substrate analog pseudouridine
-
in complex with the products, ribose 1-phosphate and uracil, at 1.8 A resolution. The biological unit is a hexamer with an alpha/beta monomeric fold. Residue His169 structurally aligns with Arg168 of the Escherichia coli uridine phosphorylase structure. A second active site residue, Lys162, is not present in previously determined uridine phosphorylase structures and interacts with O2 of uracil
Q5XA29
crystallized at 18C using the oil-microbatch method with PEG 4000 as a precipitant. A native data set is collected to 1.8 A resolution using synchrotron radiation. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 58.83, b = 76.23, c = 103.86 A, beta = 91.3; crystals: monoclinic with dimensions of 0.13 * 0.13 * 0.07 mm, X-ray diffraction to 1.8 using synchrotron radiation at -173C, belong to space group P2(1), asymmetric unit contains dimer with local pseudo-twofold symmetry, unit-cell parameters: a: 58.83, b: 76.23, c: 103.86, beta: 91.3, oil-microbatch method: 1 week at 18C, drop: 0.5 microlitre precipitant solution (27.5% (w/v) PEG 4000 in 100 mM HEPES-NaOH pH 7.5, 10 mM CaCl2) + 0.5 microlitre protein solution (21.7 mg/ml in 20 mM Tris-HCl pH8, 200 NaCl)
Q5SHF9, -
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
60
-
-
stable below
60
-
-
60 min, stable
70
-
-
half-life: 25 min in 20 mM potassium phosphate, 15.1 h in 20 mM potassium phosphate and 1 mM 5-methyluridine
70
-
-
half-life 1.6 h
70
-
Q72HS4
half-life 1.6 h
80
-
-
1 h, 20 mM potassium phosphate, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
repeated cycles of freezing and thawing inactivate the enzyme if phosphate is absent
-
the enzyme is quite labile during the course of purification, dithiothreitol and glycerol are required for stability
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, in presence of phosphate, glycerol and dithiothreitol, stable for 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant enzymes
-
harvested with 5 mM 2-mercaptoethanol and 500 mM NaCl in Tris-HCl pH 8, extract desalted with HiPrep 2610, recombinant protein eluted with 0-300 mM NaCl from Super Q Toyopearl 650M column, eluted from Bio-Scale CHT-20-1 column with 10-100 mM NaCl in phosphate-NaOH pH 7, concentrated by ultracentrifugation with 10 kDa cut-off, eluted from HiLoad 16/60 Superdex 200 prep-grade column with 200 mM NaCl in Tris-HCl pH 8, concentrated by ultracentrifugation to 21.7 mg/ml, stored at -70 C
Q5SHF9, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression of enzymes with uridine and thymidine phosphorylase activities in strain BL21(DE3)
-
expression in Escherichia coli
-
expression in Escherichia coli
-
expression in Escherichia coli
-
expression in Escherichia coli
Q72HS4
pET-11a for expression in Escherichia coli BL21 (DE3)
Q5SHF9, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
H169A
Q5XA29
about 10% of wild-type activity
H169D
Q5XA29
about 1% of wild-type activity
H169N
Q5XA29
about 4% of wild-type activity
K162A
Q5XA29
complete loss of activity
V220D
Q5XA29
almost complete loss of activity
V220E
Q5XA29
less than 0.5% of wild-type activity
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
overexpression of enzyme in Escherichia coli, medium optimization and presence of uridine and 5-fluorouracil results in conversion of more than 90% of uridine to 5-fluorouridine by the cells
synthesis
Enterobacter aerogenes CMCC 45103
-
overexpression of enzyme in Escherichia coli, medium optimization and presence of uridine and 5-fluorouracil results in conversion of more than 90% of uridine to 5-fluorouridine by the cells
-
medicine
-
the cytostatic and antiviral activity of pyrimidine nucleoside analogues is markedly decreased by a Mycoplasma hyorhinis infection and the phosphorolytic activity of the mycoplasmas is responsible for this
synthesis
-
production of ribavirin, which is an antiviral drug