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Information on EC 2.4.2.1 - purine-nucleoside phosphorylase and Organism(s) Bos taurus and UniProt Accession P55859
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2.4.2.1 purine-nucleoside phosphorylaseIUBMB Comments
Specificity not completely determined. Can also catalyse ribosyltransferase reactions of the type catalysed by EC 2.4.2.5, nucleoside ribosyltransferase.
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Word Map
- 2.4.2.1
- p-nitrophenol
- deaminase
- polynucleotide
- lymphocyte
- hypoxanthine
-
phosphorolysis
- rnase
-
salvage
- guanine
-
paraneoplastic
- t-cells
- pemphigus
- immunodeficiency
-
pincer
- deoxyguanosine
- exoribonuclease
- phosphoribosyltransferase
- polyneuropathy
- 5'-nucleotidase
-
practitioner
- dgtp
- nurse
-
degradosome
-
hypoxanthine-guanine
- parathion
- blister
-
bullous
-
mucocutaneous
- ribonucleosides
-
desmogleins
- deoxynucleosides
- deoxycytidine
- hgprt
-
phrenic
- castleman
- deoxyadenosine
- 2'-deoxyguanosine
-
metal-ligand
- obliterans
- pneumoperitoneum
- desmoplakins
- pemphigoid
-
square-planar
-
photocatalysts
- lesch-nyhan
-
iridium
-
photocatalytic
- acantholysis
- medicine
-
exonucleolytic
- p-aminophenol
- drug development
- analysis
- pharmacology
- synthesis
The taxonomic range for the selected organisms is: Bos taurus
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
pnp, purine nucleoside phosphorylase, pnpase, pfpnp, purine-nucleoside phosphorylase, adenosine phosphorylase, hspnp, hppnp, tvpnp, inosine phosphorylase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
adenosine phosphorylase
-
-
-
-
inosine phosphorylase
-
-
-
-
inosine-guanosine phosphorylase
-
-
-
-
nucleotide phosphatase (2.4.2.1)
-
-
-
-
phosphorylase, purine nucleoside
-
-
-
-
PNPase
-
-
-
-
Pu-NPase
-
-
-
-
PUNP
-
-
-
-
PUNPI
-
-
-
-
PUNPII
-
-
-
-
purine deoxynucleoside phosphorylase
-
-
-
-
purine deoxyribonucleoside phosphorylase
-
-
-
-
purine nucleoside phosphorylase
-
-
-
-
purine ribonucleoside phosphorylase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purine ribonucleoside + phosphate = purine + alpha-D-ribose 1-phosphate
the trimeric PNPs show that there is no acidic residue in the vicinity of the purine ring N7, only the side-chain of Asn243 is found there. The molecular mechanism of catalysis of trimeric PNPs involves either protonation of the purine ring N7, or a negatively charged purine intermediate stabilized by hydrogen bonds of purine N(7) with Asn243, or of the purine N1H with Glu201. Ordered water molecules provide a proton transfer bridge to O6 and N7 and permit reversible formation of these hydrogen bonds. The alternative mechanism assumes a negatively charged purine ring in the transition state stabilized by a hydrogen bond from Asn243 to purine ring N7. Key catalytic role of Glu201
purine ribonucleoside + phosphate = purine + alpha-D-ribose 1-phosphate
Theorell-Chance mechanism
-
purine ribonucleoside + phosphate = purine + alpha-D-ribose 1-phosphate
rapid equilibrium random bi-bi mechanism with formation of abortive complexes
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pentosyl group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-, -, -, -, -, -, -, -, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
purine-nucleoside:phosphate ribosyltransferase
Specificity not completely determined. Can also catalyse ribosyltransferase reactions of the type catalysed by EC 2.4.2.5, nucleoside ribosyltransferase.
CAS REGISTRY NUMBER
COMMENTARY
9030-21-1
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N7-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N8-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N9-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2-amino-6-mercapto-7-methylpurine ribonucleoside + alpha-D-ribose 1-phosphate
?
-
-
-
?
3-(beta-D-ribofuranosyl)hyopxanthine + alpha-D-ribose 1-phosphate
?
-
-
-
?
6-thioxanthine + alpha-D-ribose 1-phosphate
6-thioxanthosine + phosphate
-
-
-
?
7-(beta-D-ribofuranosyl)hypoxanthine + alpha-D-ribose 1-phosphate
?
-
-
-
?
7-methyladenosine + phosphate
7-methyladenine + alpha-D-ribose 1-phosphate
-
-
-
?
7-methylinosine + phosphate
7-methylhypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
?
8-aminoguanine + alpha-D-ribose 1-phosphate
8-aminoguanosine + phosphate
-
-
-
?
8-azaguanine + alpha-D-ribose 1-phosphate
8-azaguanine N7-D-riboside + phosphate
-
-
-
r
8-azaguanine + alpha-D-ribose 1-phosphate
8-azaguanosine + phosphate
-
-
-
?
8-azahypoxanthine + alpha-D-ribose 1-phosphate
8-azainosine + phosphate
-
-
?
deoxyguanosine + phosphate
guanine + alpha-D-deoxyribose 1-phosphate
-
-
-
?
deoxyinosine + phosphate
hypoxanthine + alpha-D-deoxyribose 1-phosphate
-
-
-
r
N7-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
the N7-riboside of 8-azaDaPur seems to be quite rapidly and specifically phosphorolysed by calf PNP. The mutation at Asn243 markedly accelerates this process
-
-
r
N8-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
wild-type enzyme PNP
-
-
r
N9-D-ribosyl-2,6-diamino-8-azapurine + phosphate
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
enzyme PNP mutant N243D
-
-
r
nicotinamide riboside + phosphate
nicotinamide + alpha-D-ribose 1-phosphate
-
-
-
?
purine deoxyribonucleoside + phosphate
purine + 2-deoxy-alpha-D-ribose 1-phosphate
-
-
-
?
purine ribonucleoside + phosphate
purine + alpha-D-ribose 1-phosphate
-
-
-
?
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N8-ribosyl-2,6-diamino-8-azapurine + N7-ribosyl-2,6-diamino-8-azapurine + phosphate
-
-
-
-
?
2-amino-6-mercapto-7-methylpurine ribonucleoside + alpha-D-ribose 1-phosphate
?
-
-
-
-
?
3-deazainosine + phosphate
alpha-D-ribose 1-phosphate + 3-deazahypoxanthine
-
-
-
-
?
7,8-dimethylguanosine + phosphate
7,8-dimethylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-butylguanosine + phosphate
7-butylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-ethylguanosine + phosphate
7-ethylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-isobutylguanosine + phosphate
7-isobutylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-isopropylguanosine + phosphate
7-isopropylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-methyl-8-aminoguanosine + phosphate
7-methyl-8-aminoguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-methyladenosine + phosphate
7-methyladenine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-methylguanosine + phosphate
7-methylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-methylinosine + phosphate
7-methylhypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
-
?
7-prolylguanosine + phosphate
7-prolylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
8-aminoguanine + alpha-D-ribose 1-phosphate
8-aminoguanosine + phosphate
-
-
-
-
?
8-azaguanine + alpha-D-ribose 1-phosphate
N9-beta-D-ribosyl-8-azaguanine + phosphate
-
-
-
-
?
8-methylguanosine + phosphate
8-methylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
deoxyguanosine + phosphate
guanine + alpha-D-deoxyribose 1-phosphate
-
-
-
-
?
deoxyinosine + phosphate
hypoxanthine + alpha-D-deoxyribose 1-phosphate
-
-
-
-
r
inosine + arsenate
hypoxanthine + ribose 1-arsenate
-
ribosyl group is nearly dissociated from the base prior to attack of the arsenate
-
?
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N7-D-ribosyl-2,6-diamino-8-azapurine + phosphate
enzyme PNP mutant D204N and mutant N243D
-
-
r
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N7-D-ribosyl-2,6-diamino-8-azapurine + phosphate
PNP mutant D204N
-
-
r
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N8-D-ribosyl-2,6-diamino-8-azapurine + phosphate
-
-
-
r
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N8-D-ribosyl-2,6-diamino-8-azapurine + phosphate
wild-type enzyme PNP
-
-
r
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N9-D-ribosyl-2,6-diamino-8-azapurine + phosphate
enzyme PNP mutant N243D
-
-
r
2,6-diamino-8-azapurine + alpha-D-ribose 1-phosphate
N9-D-ribosyl-2,6-diamino-8-azapurine + phosphate
PNP mutant D204N
-
-
r
7-methylguanosine + phosphate
7-methylguanine + alpha-D-ribose 1-phosphate
-
-
-
?
7-methylguanosine + phosphate
7-methylguanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
inosine + phosphate
hypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
?
inosine + phosphate
hypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
?
inosine + phosphate
hypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
r
8-azaguanine + alpha-D-ribose 1-phosphate
8-azaguanosine + phosphate
-
-
-
-
?
guanosine + phosphate
guanine + alpha-D-ribose 1-phosphate
-
-
-
-
?
inosine + phosphate
hypoxanthine + alpha-D-ribose 1-phosphate
-
-
-
-
?
additional information
?
-
N7-D-ribosyl-2,6-diamino-8-azapurine is a good fluorogenic substrate for mammalian forms of PNP, while the N8-riboside is selective to the Escherichia coli enzyme. The altered specificity of the PNP mutants towards some of the purine analogues can be related to differences in binding modes of 8-azapurine bases in the PNP active site, including binding in the upside-down position (rotation by 180 degree when compared with standard binding mode). Ribosylation of 2,6-diamino-8-azapurine leads to at least three fluorescent ribosides. The N9-riboside is very highly fluorescent, but its fluorescence is generally similar, in terms of lambdamax and decay time, to that of the free base. By contrast, fluorescence of N7- and N8-ribosides is red-shifted by about 70 nm and thanks to this shift their cleavage via phosphorolysis (or hydrolysis) leads to a high fluorogenic effect at about 360 nm, which can be analytically useful
-
-
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
purine deoxyribonucleoside + phosphate
purine + 2-deoxy-alpha-D-ribose 1-phosphate
-
-
-
?
purine ribonucleoside + phosphate
purine + alpha-D-ribose 1-phosphate
-
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(3S)-3-(2-amino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-(3-chlorophenyl)propanoic acid
IC50: 0.000036 mM
2,6-diamino-7-(5-mercapto-3,3-dimethylpentyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00005 mM
2,6-diamino-7-(cyclohexylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.0066 mM
2,8-diamino-3-(2-thienylmethyl)-3,9-dihydro-6H-purin-6-one
IC50: 0.001758 mM
2,8-diamino-9-(1,3-thiazol-4-ylmethyl)-1,9-dihydro-6H-purin-6-one
IC50: 0.001043 mM
2,8-diamino-9-(2-thienylmethyl)-1,9-dihydro-6H-purin-6-one
IC50: 0.000497 mM
2,8-diamino-9-(pyridin-2-ylmethyl)-1,9-dihydro-6H-purin-6-one
IC50: 0.001971 mM
2,8-diamino-9-[(4-methyl-2-thienyl)methyl]-1,9-dihydro-6H-purin-6-one
IC50: 0.000362 mM
2,8-diamino-9-[4-(1H-imidazol-1-yl)benzyl]-1,9-dihydro-6H-purin-6-one
IC50: 0.000852 mM
2,8-diamino-9-[[1-(heptyloxy)-2-hydroxyethoxy]methyl]-1,9-dihydro-6H-purin-6-one
IC50: 0.00152 mM
2-(3,4-dichlorophenyl)-5-thioxo-5,6-dihydro-1H-pyrazolo[4,3-d]pyrimidine-3,7(2H,4H)-dione
IC50: 0.00703 mM
2-amino-7-(1,2-dithian-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000018 mM
2-amino-7-(2,3,5-trichlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00024 mM
2-amino-7-(2-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00012 mM
2-amino-7-(2-furylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000083 mM
2-amino-7-(2-hydroxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00027 mM
2-amino-7-(2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000021 mM
2-amino-7-(2-thienylmethyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one
IC50: 0.000674 mM
2-amino-7-(3,4-dichlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000017 mM
2-amino-7-(3-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00002 mM
2-amino-7-(3-fluorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000024 mM
2-amino-7-(3-hydroxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00007 mM
2-amino-7-(3-methoxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000082 mM
2-amino-7-(3-methylbenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000057mM
2-amino-7-(3-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000025 mM
2-amino-7-(4-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000025 mM
2-amino-7-(4-iodobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000023 mM
2-amino-7-(biphenyl-4-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000546 mM
2-amino-7-(cycloheptylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00003 mM
2-amino-7-(cyclohexylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000047 mM
2-amino-7-(cyclopentylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000029 mM
2-amino-7-(piperidin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.001 mM
2-amino-7-(pyridin-2-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000015 mM
2-amino-7-(pyridin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000025 mM
2-amino-7-(pyridin-4-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000064 mM
2-amino-7-(tetrahydro-2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000011 mM
2-amino-7-(tetrahydrofuran-2-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00007 mM
2-amino-7-benzyl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000051 mM
2-amino-7-cyclohex-1-en-1-yl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.0019 mM
2-amino-7-phenoxy-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000042 mM
2-amino-7-[(3-methylcyclohexyl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000025 mM
2-amino-7-[(3-tert-butylcyclohexyl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000025 mM
2-amino-7-[(4-chloropyridin-3-yl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000135 mM
2-amino-7-[3-(trifluoromethyl)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000036 mM
2-amino-7-[[(1R,3S,5S,7S)-4-methyl-2-adamantyl]methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00009 mM
2-amino-9-(2-thienylmethyl)-8-thioxo-1,7,8,9-tetrahydro-6H-purin-6-one
IC50: 0.002866 mM
2-phenyl-5,7-dithioxo-1,2,4,5,6,7-hexahydro-3H-pyrazolo[4,3-d]pyrimidin-3-one
IC50: 0.02284 mM
3-(2-amino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-phenylpropanoic acid
IC50: 0.000047 mM
3-(3-chlorophenyl)-3-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)propanenitrile
IC50: 0.000011 mM
3-(3-chlorophenyl)-3-(4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)propanenitrile
IC50: 0.00001 mM
3-(4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-pyridin-3-ylpropanenitrile
IC50: 0.000039 mM
4-imino-7-methyl-1,2,3,4-tetrahydropyrazolo [1,5-a] [1,3,5] triazin-2-one
-
5-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3,3-dimethylpentane-1-sulfonamide
IC50: 0.0001 mM
5-amino-3-(2-thienylmethyl)-3,6-dihydro-7H-[1,2,3]triazolo[4,5-d]pyrimidin-7-one
IC50: 0.001839 mM
6-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-4,4-dimethylhexanamide
IC50: 0.2 mM
6-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-4,4-dimethylhexanenitrile
IC50: 0.00026 mM
6-amino-7-(2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00016 mM
7-(3-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000028 mM
7-(cyclopentylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000029 mM
7-(pyridin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00004 mM
7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.00019 mM
SerMe-immucillin-H
SerMe-ImmH, uses achiral dihydroxyaminoalcohol seramide as the ribocation mimic
(+/-)-cis-1,1-difluoro-2-(tetrahydro-3-piranozyl)ethylphosphonic acid
-
with (hypoxanthine-9-yl)methyl aglycone, i.e. Yokomatsu compound
(+/-)-cis-1,1-difluoro-2-(tetrahydro-3-piranyl)ethylphosphonic acid with (hypoxanthine-9-yl)methyl aglycone
-
multisubstrate analogue inhibitor
2-amino-6-mercapto-7-methylpurine ribonucleoside
-
in absence of phosphate the enzyme catalyzes a slow hydrolysis, which is accompanied by inactivation of the enzyme
7-ketopyrazolo[4,3-d]pyrimidine
-
inhibits phosphorolysis of 7-methylguanosine uncompetitively, inhibits synthesis of 8-azaguanosine competitively
7KPP
-
non-substrate inhibitor of both the phosphorolytic and reverse synthetic pathways, inhibits synthesis of 8-azaguanosine from 8-azaguanine competitively
9-(2'-benzyl-5',5'-difluoro-5'-phosphonopentyl)guanine
-
multisubstrate analogue inhibitor
9-(5',5'-difluoro-5'-phosphonobutyl)-9-deazaguanine
-
multisubstrate analogue inhibitor
9-(5',5'-difluoro-5'-phosphonoheptyl)-9-deazaguanine
-
multisubstrate analogue inhibitor
cis-1,1-difluoro-2-(tetrahydro-3-piranozyl)ethylphosphonic acid
-
with (hypoxanthine-9-yl)methyl aglycone
additional information
-
inhibition by 9-cycloaliphatic methyl and 9-saturated heterocyclic methyl-9-deazapurines
-
2-amino-7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000124 mM
2-amino-7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
IC50: 0.000147 mM
9-(2-phosphonylmethoxyethyl)-8-azaguanine
-
inhibits phosphorolytic and synthetic reaction, competitive
9-(2-phosphonylmethoxyethyl)-8-azaguanine
-
competitive versus 8-azaguanine and alpha-D-ribose 1-phosphate
9-(5',5'-difluoro-5'-phosphonopentyl)-9-deazaguanine
-
multisubstrate analogue inhibitor. Upon titration of the recombinant enzyme, endothermic as well as exothermic signals are obtained. Part of the recombinant enzyme forms a complex with its product, hypoxanthine, although hypoxanthine was not present at any purification stage except for its natural occurrence in Escherichia coli cells. Binding of hypoxanthine is accompanied with a large negative change of the free enthalpy, and therefore the replacement of hypoxanthine by 9-(5',5'-difluoro-5'-phosphonopentyl)-9-deazaguanine yields a positive heat signal
formycin B
-
weak, uncompetitive inhibitor. Formycin B forms a weakly fluorescent complex with the enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.05
N7-D-ribosyl-2,6-diamino-8-azapurine
about, pH 6.5, 25°C, recombinant mutant N243D
0.052
N7-D-ribosyl-2,6-diamino-8-azapurine
pH 6.5, 25°C, recombinant wild-type enzyme
0.035
2,6-diamino-8-azapurine
-
mutant enzyme N243D, pH and temperature not specified in the publication
0.06
2,6-diamino-8-azapurine
-
wild type enzyme, pH and temperature not specified in the publication
0.0168
7-methylguanosine
-
wild-type, pH not specified in the publication, temperature not specified in the publication
0.05
7-methylguanosine
-
mutant F159W, pH not specified in the publication, temperature not specified in the publication
0.07
7-methylguanosine
-
mutant F200W, pH not specified in the publication, temperature not specified in the publication
0.09
8-Azaguanine
-
wild type enzyme, pH and temperature not specified in the publication
0.1
8-Azaguanine
-
Km above 0.1 mM, mutant enzyme N243D, pH and temperature not specified in the publication
0.0134
Inosine
-
wild-type, pH not specified in the publication, temperature not specified in the publication
0.245
Inosine
-
mutant F159W, pH not specified in the publication, temperature not specified in the publication
0.326
Inosine
-
mutant F200W, pH not specified in the publication, temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000033
9-(5',5'-difluoro-5'-phosphonopentyl)-9-deazaguanine
pH 7.0, 25°C, in the presence of 1 mM phosphate
0.000000023
DADMe-immucillin-H
pH and temperature not specified in the publication
0.000001
(+/-)-cis-1,1-difluoro-2-(tetrahydro-3-piranozyl)ethylphosphonic acid
-
approximate value, with (hypoxanthine-9-yl)methyl aglycone, i.e. Yokomatsu compound
0.0000112
(+/-)-cis-1,1-difluoro-2-(tetrahydro-3-piranyl)ethylphosphonic acid with (hypoxanthine-9-yl)methyl aglycone
-
pH 7.0, 25°C
0.0069
9-(5',5'-difluoro-5'-phosphonopentyl)-9-deazaguanine
-
wild-type, pH not specified in the publication, temperature not specified in the publication
0.063
cis-1,1-difluoro-2-(tetrahydro-3-piranozyl)ethylphosphonic acid
-
wild-type, pH not specified in the publication, temperature not specified in the publication
0.0313
guanosine
-
with guanine as variable substrate and 0.4 mM ribose 1-phosphate
0.0021
1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamidine
-
reaction with 7-methylguanosine
0.0062
1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamidine
-
reaction with inosine
0.002
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 7.3, concentration of the substrate 8-azaguanosine is 0.005-0.05 mM
0.0028 - 0.0035
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 6.5, concentration of the substrate 8-azaguanosine is 0.05-0.25 mM
0.0036
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 7.9, concentration of the substrate 8-azaguanosine is 0.005-0.05 mM
0.0037
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 8.2, concentration of the substrate 8-azaguanosine is 0.005-0.05 mM
0.0042
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 8.2, concentration of the substrate 8-azaguanosine is 0.005-0.05 mM
0.038
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 6.0, concentration of the substrate 7-methylguanosine is 0.032 mM
0.054
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 7.0, concentration of the substrate 7-methylguanosine is 0.03 mM
0.073
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 7.0, concentration of the substrate 7-methylguanosine is 0.122 mM
0.093
7-ketopyrazolo[4,3-d]pyrimidine
-
pH 8.0, concentration of the substrate 7-methylguanosine is 0.032 mM
0.32
alpha-D-ribose 1-phosphate
-
with phosphate as variable substrate and 0.2 mM guanosine
0.36
alpha-D-ribose 1-phosphate
-
with guanosine as variable substrate and 20 mM phosphate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000036
(3S)-3-(2-amino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-(3-chlorophenyl)propanoic acid
Bos taurus
IC50: 0.000036 mM
0.00005
2,6-diamino-7-(5-mercapto-3,3-dimethylpentyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00005 mM
0.0066
2,6-diamino-7-(cyclohexylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.0066 mM
0.001758
2,8-diamino-3-(2-thienylmethyl)-3,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.001758 mM
0.001043
2,8-diamino-9-(1,3-thiazol-4-ylmethyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.001043 mM
0.000497
2,8-diamino-9-(2-thienylmethyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.000497 mM
0.00417
2,8-diamino-9-(4-fluorobenzyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.00417 mM
0.001971
2,8-diamino-9-(pyridin-2-ylmethyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.001971 mM
0.000362
2,8-diamino-9-[(4-methyl-2-thienyl)methyl]-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.000362 mM
0.000852
2,8-diamino-9-[4-(1H-imidazol-1-yl)benzyl]-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.000852 mM
0.00152
2,8-diamino-9-[[1-(heptyloxy)-2-hydroxyethoxy]methyl]-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.00152 mM
0.00703
2-(3,4-dichlorophenyl)-5-thioxo-5,6-dihydro-1H-pyrazolo[4,3-d]pyrimidine-3,7(2H,4H)-dione
Bos taurus
IC50: 0.00703 mM
0.000018
2-amino-7-(1,2-dithian-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000018 mM
0.00024
2-amino-7-(2,3,5-trichlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00024 mM
0.00012
2-amino-7-(2-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00012 mM
0.000083
2-amino-7-(2-furylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000083 mM
0.00027
2-amino-7-(2-hydroxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00027 mM
0.000021
2-amino-7-(2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000021 mM
0.000674
2-amino-7-(2-thienylmethyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one
Bos taurus
IC50: 0.000674 mM
0.000017
2-amino-7-(3,4-dichlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000017 mM
0.00002
2-amino-7-(3-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00002 mM
0.000024
2-amino-7-(3-fluorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000024 mM
0.00007
2-amino-7-(3-hydroxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00007 mM
0.000082
2-amino-7-(3-methoxybenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000082 mM
0.000057
2-amino-7-(3-methylbenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000057mM
0.000025
2-amino-7-(3-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000025 mM
0.000025
2-amino-7-(4-chlorobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000025 mM
0.000023
2-amino-7-(4-iodobenzyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000023 mM
0.000546
2-amino-7-(biphenyl-4-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000546 mM
0.00003
2-amino-7-(cycloheptylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00003 mM
0.000047
2-amino-7-(cyclohexylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000047 mM
0.000029
2-amino-7-(cyclopentylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000029 mM
0.001
2-amino-7-(piperidin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.001 mM
0.000015
2-amino-7-(pyridin-2-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000015 mM
0.000025
2-amino-7-(pyridin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000025 mM
0.000064
2-amino-7-(pyridin-4-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000064 mM
0.000011
2-amino-7-(tetrahydro-2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000011 mM
0.00007
2-amino-7-(tetrahydrofuran-2-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00007 mM
0.000051
2-amino-7-benzyl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000051 mM
0.0019
2-amino-7-cyclohex-1-en-1-yl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.0019 mM
0.000042
2-amino-7-phenoxy-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000042 mM
0.000025
2-amino-7-[(3-methylcyclohexyl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000025 mM
0.000025
2-amino-7-[(3-tert-butylcyclohexyl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000025 mM
0.000135
2-amino-7-[(4-chloropyridin-3-yl)methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000135 mM
0.000124 - 0.000147
2-amino-7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
0.000036
2-amino-7-[3-(trifluoromethyl)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000036 mM
0.00009
2-amino-7-[[(1R,3S,5S,7S)-4-methyl-2-adamantyl]methyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00009 mM
0.002866
2-amino-9-(2-thienylmethyl)-8-thioxo-1,7,8,9-tetrahydro-6H-purin-6-one
Bos taurus
IC50: 0.002866 mM
0.02284
2-phenyl-5,7-dithioxo-1,2,4,5,6,7-hexahydro-3H-pyrazolo[4,3-d]pyrimidin-3-one
Bos taurus
IC50: 0.02284 mM
0.000047
3-(2-amino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-phenylpropanoic acid
Bos taurus
IC50: 0.000047 mM
0.000011
3-(3-chlorophenyl)-3-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)propanenitrile
Bos taurus
IC50: 0.000011 mM
0.00001
3-(3-chlorophenyl)-3-(4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)propanenitrile
Bos taurus
IC50: 0.00001 mM
0.000039
3-(4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3-pyridin-3-ylpropanenitrile
Bos taurus
IC50: 0.000039 mM
0.0001
5-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-3,3-dimethylpentane-1-sulfonamide
Bos taurus
IC50: 0.0001 mM
0.001839
5-amino-3-(2-thienylmethyl)-3,6-dihydro-7H-[1,2,3]triazolo[4,5-d]pyrimidin-7-one
Bos taurus
IC50: 0.001839 mM
0.2
6-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-4,4-dimethylhexanamide
Bos taurus
IC50: 0.2 mM
0.00026
6-(2,6-diamino-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)-4,4-dimethylhexanenitrile
Bos taurus
IC50: 0.00026 mM
0.00016
6-amino-7-(2-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00016 mM
0.000028
7-(3-thienylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000028 mM
0.000029
7-(cyclopentylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000029 mM
0.00004
7-(pyridin-3-ylmethyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00004 mM
0.000035
7-benzyl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000035 mM
0.00019
7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.00019 mM
0.000092
2,8-diamino-9-(2-furylmethyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.000092 mM
0.00271
2,8-diamino-9-(2-furylmethyl)-1,9-dihydro-6H-purin-6-one
Bos taurus
IC50: 0.00271 mM
0.000124
2-amino-7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000124 mM
0.000147
2-amino-7-[3-(benzyloxy)benzyl]-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one
Bos taurus
IC50: 0.000147 mM
0.098
7-ketopyrazolo[4,3-d]pyrimidine
Bos taurus
-
pH 7.0, concentration of the substrate 7-methylguanosine is 0.03 mM
0.098
7-ketopyrazolo[4,3-d]pyrimidine
Bos taurus
-
pH 7.0, concentration of the substrate 7-methylguanosine is 0.122 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
substrate specificity of trimeric and hexameric PNPs may be changed by mutations of the crucial active site amino acids, namely Asp in hexameric PNPs and Asn in trimeric PNPs
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PNPH_BOVIN
289
0
32037
Swiss-Prot
other Location (Reliability: 1)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotrimer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
high-resolution structure may serve for design of inhibitors with potential pharmacological application
binary complex of the trimeric enzyme with the potent ground-state analog inhibitor 9-(5,5-difluoro-5-phosphonopentyl)guanine crystallized in the cubic space group P2(1)3 with unit-cell parameter a = 93.183 A
-
calorimetric titration of recombinant calf PNP complexed with immucillin H. Stoichiometry shows three immucillin molecules per enzyme trimer, enzyme does not show negative cooperativity. One-third-of-the-sites binding does not occur for trimeric enzyme
-
hanging drop method, purine nucleoside phosphorylase complexed with substrates and substrate analogues
-
X-ray crystal structure for purine nucleoside phosphorylase with bound 9-deazainosine and inorganic sulfate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
F159W
the mutant shows reduced activity and stability compared to the wild type enzyme
F200W
the mutant shows reduced activity and stability compared to the wild type enzyme
N123L/R210Q
mutation of bovine enzyme toward human enzyme. Steady-state kinetic studies indicate unchanged catalytic activity, the mutant enzyme has higher affinity for inhibitors that are mimics of a late dissociative transition state
N243D
V39T/N123L/R210Q
mutation of bovine enzyme toward human enzyme. Steady-state kinetic studies indicate unchanged catalytic activity, while presteady-state studies indicate that the chemical step is slower in the triple mutant. The mutant enzyme has higher affinity for inhibitors that are mimics of a late dissociative transition state. Mutant displays a highly dissociative SN1 transition state with low bond order to the leaving group, a transition state different from the native enzyme, with significant nucleophilic participation at the transition state
L22E/H104R
-
mutation of human toward bovine purine nucleoside phosphorylase. Mutant is similar to human purine nucleoside phosphorylase in steady-state kinetic properties. Inhibitor DADMe-immucillin-G is an excellent mimic of the transition states for both human purine nucleoside phosphorylase and the mutant with a preference for the mutant. Thermodynamic parameters establish the mutant to be catalytically more efficient than the parent enzyme and reveal differences in the entropic component of catalysis
N243D
site-directed mutagenesis, the mutant exhibits altered substrate specificity compared to wild-type
N243D
site-directed mutagenesis, the mutation in trimeric PNP changes the substrate specificity, making 6-aminopurine nucleosides good substrates
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
strong susceptibility to photooxidation in presence of methylene blue. Maximal photoinactivation at pH 8.0, 85% loss of activity after only 1 min of irradiation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 10 mM sodium phosphate buffer, pH 7.0, stable for at least 4 months provided a weekly dialysis with 2 mM DTT is performed
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli BL21 (DE3). The enzyme shows high activity and behavior similar to the protein isolated from mammalian tissue. Thus it can fully replace the non-recombinant PNP
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
differences in specificity between homotrimeric (including human enzyme) and homohexameric PNPs, including various pathogenic organisms, make them interesting potential drug targets
medicine
synthesis
enzymatic ribosylation of fluorescent 8-azapurine derivatives, like 8-azaguanine and 2,6-diamino-8-azapurine, with purine-nucleoside phosphorylase (PNP) as a catalyst, leads to N9, N8, and N7-ribosides. The final proportion of the products may be modulated by point mutations in the enzyme active site. As an example, ribosylation of the latter substrate by wild-type calf PNP gives N7- and N8-ribosides, while the N243D mutant directs the ribosyl substitution at N9- and N7-positions. The same mutant allows synthesis of the fluorescent N7-D-ribosyl-8-azaguanine. The N7- and N8-ribosides of the 8-azapurines can be analytically useful, as illustrated by N7-D-ribosyl-2,6-diamino-8-azapurine, which is a good fluorogenic substrate for mammalian forms of PNP, while the N8-riboside is selective to the Escherichia coli enzyme
medicine
high-resolution structure may serve for design of inhibitors with potential pharmacological application
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Agarwal, R.P.; Parks, R.E.
Purine nucleoside phosphorylase from human erythrocytes. IV. Crystallization and some properties
J. Biol. Chem.
244
644-647
1969
Bos taurus, Homo sapiens
Parks, R.E.; Agarwal, R.P.
Purine nucleoside phosphorylase
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
483-514
1972
Klebsiella aerogenes, Aeromonas hydrophila, Brevibacillus brevis, Bacillus cereus, Bacillus subtilis, Bacillus licheniformis, Bacterium cadaveris, Bos taurus, Canis lupus familiaris, Gallus gallus, Columba livia, Clavibacter michiganensis subsp. sepedonicus, Oryctolagus cuniculus, Escherichia coli, Equus caballus, Pectobacterium carotovorum, Felis catus, Homo sapiens, Lactobacillus leichmannii, Leucisus rusticus, Macaca mulatta, Micrococcus luteus, Papio hamadryas, Proteus vulgaris, Shewanella putrefaciens, Rattus norvegicus, salmon, Salmonella enterica subsp. enterica serovar Enteritidis, Sus scrofa
-
Moyer, T.P.; Fischer, A.G.
Purification and characterization of a purine-nucleoside phosphorylase from bovine thyroid
Arch. Biochem. Biophys.
174
622-629
1976
Bos taurus
Barsacchi, D.; Cappiello, M.; Tozzi, M.G.; Del Corso, A.; Peccatori, M.; Camici, M.; Ipata, P.L.; Mura, U.
Purine nucleoside phosphorylase from bovine lens: purification and properties
Biochim. Biophys. Acta
1160
163-170
1992
Bos taurus
Lewis, A.S.; Glantz, M.D.
Bovine brain purine-nucleoside phosphorylase purification, characterization, and catalytic mechanism
Biochemistry
15
4451-4457
1976
Bos taurus
Bzowska, A.; Kulikowska, E.; Darzynkiewicz, E.; Shugar, D.
Purine nucleoside phosphorylase. Structure-activity relationships for substrate and inhibitor properties of N-1-, N-7-, and C-8-substituted analogues; differentiation of mammalian and bacterial enzymes with N-1-methylinosine and guanosine
J. Biol. Chem.
263
9212-9217
1988
Bos taurus, Escherichia coli
Kulikowska, E.; Bzowska, A.; Wierzchowski, J.; Shugar, D.
Properties of two unusual, and fluorescent, substrates of purine-nucleoside phosphorylase: 7-methylguanosine and 7-methylinosine
Biochim. Biophys. Acta
874
355-363
1986
Bos taurus
Bzowska, A.; Kulikowska, E.; Shugar, D.
Properties of purine nucleoside phosphorylase (PNP) of mammalian and bacterial origin
Z. Naturforsch. C
45c
59-70
1990
Bos taurus, Escherichia coli, Homo sapiens
-
Mao, C.; Cook, W.J.; Zhou, M.; Federov, A.A.; Almo, S.C.; Ealick, S.E.
Calf spleen purine nucleoside phosphorylase complexed with substrates and substrate analogues
Biochemistry
36
7135-7146
1998
Bos taurus
-
Bzowska, A.; Kulikowska, E.; Poopeiko, N.E.; Shugar, D.
Kinetics of phosphorolysis of 3-(beta-D-ribofuranosyl)adenine and 3-(beta-D-ribofuranosyl)hypoxanthine, non-conventional substrates of purine-nucleoside phosphorylase
Eur. J. Biochem.
239
229-234
1996
Bos taurus, Escherichia coli
Kline, P.C.; Schramm, V.L.
Purine nucleoside phosphorylase. Catalytic mechanism and transition-state analysis of the arsenolysis reaction
Biochemistry
32
13212-13219
1993
Bos taurus
Koeller, G.; Luic, M.; Shugar, D.; Saenger, W.; Bzowska, A.
Crystal structure of calf spleen purine nucleoside phosphorylase in a complex with hypoxanthine at 2.15 A resolution
J. Mol. Biol.
265
202-216
1997
Bos taurus (P55859)
Stoychev, G.; Kierdaszuk, B.; Shugar, D.
Xanthosine and xanthine. Substrate properties with purine nucleoside phosphorylases, and relevance to other enzyme systems
Eur. J. Biochem.
269
4048-4057
2002
Homo sapiens, Bos taurus (P55859)
Montgomery, J.A.
Purine nucleoside phosphorylase: a target for drug design
Med. Res. Rev.
13
209-228
1993
Bos taurus, Homo sapiens
Cheng, J.; Farutin, V.; Wu, Z.; Jacob-Mosier, G.; Riley, B.; Hakimi, R.; Cordes, E.H.
Purine nucleoside phosphorylase-catalyzed, phosphate-independent hydrolysis of 2-amino-6-mercapto-7-methylpurine ribonucleoside
Bioorg. Chem.
27
307-325
1999
Bos taurus
-
Bzowska, A.; Kulikowska, E.; Shugar, D.
Purine nucleoside phosphorylases: properties, functions, and clinical aspects
Pharmacol. Ther.
88
349-425
2000
Acholeplasma laidlawii, Klebsiella aerogenes, Geobacillus stearothermophilus, Bacillus cereus, Cellulomonas sp., Pectobacterium carotovorum, Fasciola hepatica, Klebsiella sp., Micrococcus luteus, Mus musculus, Plasmodium falciparum, Plasmodium lophurae, Proteus vulgaris, Rattus norvegicus, Salmonella enterica subsp. enterica serovar Typhimurium, Serratia marcescens, Saccharolobus solfataricus, Trypanosoma brucei, Trypanosoma cruzi, Homo sapiens (P00491), Homo sapiens, Escherichia coli (P0ABP8), Bacillus subtilis (P46354), Bos taurus (P55859), Saccharomyces cerevisiae (Q05788)
Luic, M.; Koellner, G.; Yokomatsu, T.; Shibuya, S.; Bzowska, A.
Calf spleen purine-nucleoside phosphorylase: crystal structure of the binary complex with a potent multisubstrate analogue inhibitor
Acta Crystallogr. Sect. D
60
1417-1424
2004
Bos taurus
Iwanow, M.; Magnowska, L.; Yokomatsu, T.; Shibuya, S.; Bzowska, A.
Interactions of potent multisubstrate analogue inhibitors with purine nucleoside phosphorylase from calf spleen--kinetic and spectrofluorimetric studies
Nucleosides Nucleotides Nucleic Acids
22
1567-1570
2003
Bos taurus
Wierzchowski, J.; Bzowska, A.; Stepniak, K.; Shugar, D.
Interactions of calf spleen purine nucleoside phosphorylase with 8-azaguanine, and a bisubstrate analogue inhibitor: implications for the reaction mechanism
Z. Naturforsch. C
59
713-725
2004
Bos taurus
Toms, A.V.; Wang, W.; Li, Y.; Ganem, B.; Ealick, S.E.
Novel multisubstrate inhibitors of mammalian purine nucleoside phosphorylase
Acta Crystallogr. Sect. D
D61
1449-1458
2005
Bos taurus (P55859), Bos taurus
Rinaldo-Matthis, A.; Wing, C.; Ghanem, M.; Deng, H.; Wu, P.; Gupta, A.; Tyler, P.C.; Evans, G.B.; Furneaux, R.H.; Almo, S.C.; Wang, C.C.; Schramm, V.L.
Inhibition and structure of Trichomonas vaginalis purine nucleoside phosphorylase with picomolar transition state analogues
Biochemistry
46
659-668
2007
Trichomonas vaginalis, Homo sapiens (P00491), Homo sapiens, Bos taurus (P55859), Plasmodium falciparum (Q8T9Z7)
Castilho, M.S.; Postigo, M.P.; de Paula, C.B.; Montanari, C.A.; Oliva, G.; Andricopulo, A.D.
Two- and three-dimensional quantitative structure-activity relationships for a series of purine nucleoside phosphorylase inhibitors
Bioorg. Med. Chem.
14
516-527
2006
Bos taurus (P55859)
Silva, R.G.; Nunes, J.E.; Canduri, F.; Borges, J.C.; Gava, L.M.; Moreno, F.B.; Basso, L.A.; Santos, D.S.
Purine nucleoside phosphorylase: a potential target for the development of drugs to treat T-cell- and apicomplexan parasite-mediated diseases
Curr. Drug Targets
8
413-422
2007
Plasmodium lophurae, Toxoplasma gondii, Homo sapiens (P00491), Homo sapiens, Bos taurus (P55859), Plasmodium falciparum (Q8T9Z7)
Wierzchowski, J.; Stepniak, K.; Bzowska, A.; Shugar, D.
Spectroscopic and kinetic studies of interactions of calf spleen purine nucleoside phosphorylase with 8-azaguanine, and its 9-(2-phosphonylmethoxyethyl) derivative
Nucleosides Nucleotides Nucleic Acids
24
459-464
2005
Bos taurus
Pereira, H.M.; Berdini, V.; Cleasby, A.; Garratt, R.C.
Crystal structure of calf spleen purine nucleoside phosphorylase complexed to a novel purine analogue
FEBS Lett.
581
5082-5086
2007
Bos taurus (P55859), Bos taurus
Wierzchowski, J.; Iwanska, B.; Bzowska, A.; Shugar, D.
Interactions of calf spleen purine nucleoside phosphorylase with formycin B and its aglycone - spectroscopic and kinetic studies
Nucleosides Nucleotides Nucleic Acids
26
849-854
2007
Bos taurus
Stepniak, K.; Girstun, A.; Wielgus-Kutrowska, B.; Staro?, K.; Bzowska, A.
Cloning, expression, purification, and some properties of calf purine nucleoside phosphorylase
Nucleosides Nucleotides Nucleic Acids
26
855-859
2007
Bos taurus
Glavas-Obrovac, L.; Suver, M.; Hikishima, S.; Yokomatsu, T.; Bzowska, A.
Inhibitory properties of nucleotides with difluoromethylenephosphonic acid as a phosphate mimic versus calf spleen purine nucleoside phosphorylase and effect of these analogues on the viability of human blood lymphocytes
Nucleosides Nucleotides Nucleic Acids
26
989-993
2007
Bos taurus
Breer, K.; Girstun, A.; Wielgus-Kutrowska, B.; Staro?, K.; Bzowska, A.
Overexpression, purification and characterization of functional calf purine nucleoside phosphorylase (PNP)
Protein Expr. Purif.
61
122-130
2008
Bos taurus (P55859), Bos taurus
Breer, K.; Wielgus-Kutrowska, B.; Girstun, A.; Staro?, K.; Hashimoto, M.; Hikishima, S.; Yokomatsu, T.; Bzowska, A.
Overexpressed proteins may act as mops removing their ligands from the host cells: a case study of calf PNP
Biochem. Biophys. Res. Commun.
391
1203-1209
2010
Bos taurus
Ghanem, M.; Li, L.; Wing, C.; Schramm, V.L.
Altered thermodynamics from remote mutations altering human toward bovine purine nucleoside phosphorylase
Biochemistry
47
2559-2564
2008
Bos taurus, Homo sapiens
Li, L.; Luo, M.; Ghanem, M.; Taylor, E.A.; Schramm, V.L.
Second-sphere amino acids contribute to transition-state structure in bovine purine nucleoside phosphorylase
Biochemistry
47
2577-2583
2008
Homo sapiens (P00491), Homo sapiens, Bos taurus (P55859), Bos taurus
Glavas-Obrovac, L.; Suver, M.; Hikishima, S.; Hashimoto, M.; Yokomatsu, T.; Magnowska, L.; Bzowska, A.
Antiproliferative activity of purine nucleoside phosphorylase multisubstrate analogue inhibitors containing difluoromethylene phosphonic acid against leukaemia and lymphoma cells
Chem. Biol. Drug Des.
75
392-399
2010
Bos taurus, Homo sapiens
Wielgus-Kutrowska, B.; Breer, K.; Hashimoto, M.; Hikishima, S.; Yokomatsu, T.; Narczyk, M.; Dyzma, A.; Girstun, A.; Staro?, K.; Bzowska, A.
Trimeric purine nucleoside phosphorylase: exploring postulated one-third-of-the-sites binding in the transition state
Bioorg. Med. Chem.
20
6758-6769
2012
Bos taurus
Wielgus-Kutrowska, B.; Modrak-Wojcik, A.; Dyzma, A.; Breer, K.; Zolkiewski, M.; Bzowska, A.
Purine nucleoside phosphorylase activity decline is linked to the decay of the trimeric form of the enzyme
Arch. Biochem. Biophys.
549
40-48
2014
Bos taurus (P55859)
Stachelska-Wierzchowska, A.; Wierzchowski, J.; Bzowska, A.; Wielgus-Kutrowska, B.
Site-selective ribosylation of fluorescent nucleobase analogs using purine-nucleoside phosphorylase as a catalyst: effects of point mutations
Molecules
21
E44
2015
Bos taurus
Wierzchowski, J.; Stachelska-Wierzchowska, A.; Wielgus-Kutrowska, B.; Bzowska, A.
1,N6-ethenoadenine and other fluorescent nucleobase analogs as substrates for purine-nucleoside phosphorylases Spectroscopic and kinetic studies
Curr. Pharm. Des.
23
6948-6966
2017
Bacillus cereus, Pectobacterium carotovorum, Thermus thermophilus, Plasmodium lophurae, Helicobacter pylori (A0A518Y5Z2), Homo sapiens (P00941), Escherichia coli (P0ABP8), Escherichia coli (P45563), Bos taurus (P55859), Cellulomonas sp. (P81989), Plasmodium falciparum (Q8I3X4)
Stachelska-Wierzchowska, A.; Wierzchowski, J.; Bzowska, A.; Wielgus-Kutrowska, B.
Site-selective ribosylation of fluorescent nucleobase analogs using purine-nucleoside phosphorylase as a catalyst effects of point mutations
Molecules
21
44
2016
Homo sapiens (P00491), Homo sapiens, Escherichia coli (P0ABP8), Escherichia coli, Bos taurus (P55859)
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