Literature summary for 2.4.2.1 extracted from

Timofeev, V.I.; Zhukhlistova, N.E.; Abramchik, Y.A.; Muravieva, T.I.; Esipov, R.S.; Kuranova, I.P.
Crystal structure of Escherichia coli purine nucleoside phosphorylase complexed with acyclovir (2018), Acta Crystallogr. Sect. F, 74, 402-409 .

Application

Application	Comment	Organism
drug development	owing to their key role in the purine salvage pathway, PNPs are attractive targets for drug design against some pathogens	Escherichia coli

Cloned(Commentary)

Cloned (Comment)	Organism
gene pnp, sequence comparisons of Escherichia coli enzyme with human and Bacillus subtilis enzymes, recombinant expression in Escherichia coli strain BL21(DE3)/pERPUPHOI	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme PNP in complex with inhibitor ACV, liquid diffusion method, mixing of 0.0035 ml of 32 mg/ml protein in 0.02 M Tris-HCl, pH 7.5, with 0.0035 ml of reservoir solution containing 25% ammonium sulfate, 0.05 M sodium citrate, pH 4.9, 0.04% sodium azide, and 5 mM acycloguanosine, and equilbration against 0.18 ml of reservoir solution, X-ray diffraction structure determination at 2.32 A resolution using the molecular replacement method. The ACV molecule is observed in two conformations and sulfate ions are located in both the nucleoside-binding and phosphate-binding pockets of the enzyme	Escherichia coli

Crystallization (Comment)

Organism

purified enzyme PNP in complex with inhibitor ACV, liquid diffusion method, mixing of 0.0035 ml of 32 mg/ml protein in 0.02 M Tris-HCl, pH 7.5, with 0.0035 ml of reservoir solution containing 25% ammonium sulfate, 0.05 M sodium citrate, pH 4.9, 0.04% sodium azide, and 5 mM acycloguanosine, and equilbration against 0.18 ml of reservoir solution, X-ray diffraction structure determination at 2.32 A resolution using the molecular replacement method. The ACV molecule is observed in two conformations and sulfate ions are located in both the nucleoside-binding and phosphate-binding pockets of the enzyme

Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
acyclovir	ACV, an acyclic derivative of the PNP substrate guanosine, an acyclic analogue of 2'-deoxyguanosine, that is used as an antiviral drug for the treatment of some human viral infections. The ACV molecule occupies the nucleoside-binding pocket in the active-site cavity, where it adopts two conformations. Sulfate ions are located in both the nucleoside-binding and phosphate-binding pockets of the enzyme. Binding structure, overview	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
adenosine + phosphate	Escherichia coli	-	adenine + alpha-D-ribose 1-phosphate	-	r
guanosine + phosphate	Escherichia coli	-	guanine + alpha-D-ribose 1-phosphate	-	r
inosine + phosphate	Escherichia coli	-	hypoxanthine + alpha-D-ribose 1-phosphate	-	r

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P0ABP8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21(DE3)/pERPUPHOI cell extract supernatant by anion exchange chromatography, ultrafiltration, and gel filtration	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
adenosine + phosphate	-	Escherichia coli	adenine + alpha-D-ribose 1-phosphate	-	r
guanosine + phosphate	-	Escherichia coli	guanine + alpha-D-ribose 1-phosphate	-	r
inosine + phosphate	-	Escherichia coli	hypoxanthine + alpha-D-ribose 1-phosphate	-	r

Subunits

Subunits	Comment	Organism
homohexamer	the hexameric molecule can be considered as a trimer of dimers, where each dimer contains two complete active sites. Each monomer consists of a central core comprising mixed beta-sheet surrounded by alpha-helices with an extended C-terminal alpha7 helix. The active sites are located within the deep cavity between the dimer subunits and comprise amino-acid residues from both subunits. The active-site cavity is predominantly lined by hydrophobic amino acids from strands beta5, beta7, beta8, beta9, and beta10 and helix alpha7. The adjacent dimer subunit contributes the side chains of His4* and Arg43* to the active site	Escherichia coli

Synonyms

Synonyms	Comment	Organism
DeoD	-	Escherichia coli
PNP	-	Escherichia coli
purine nucleoside phosphorylase	-	Escherichia coli

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the type I PNP family of hexameric enzymes	Escherichia coli
metabolism	PNPs play a key role in the purine salvage pathway	Escherichia coli
physiological function	the enzyme catalyzes the reversible phosphorolysis of purine ribonucleosides	Escherichia coli