Dr Larissa Balakireva, CEO & Founder of NovoCIB, was awarded with the Trophy of
"Femmes en Or 2011, Femme de l'Innovation"
in September 2011

Ref. #E-Nov6
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50 mUnits € 305.00
150 mUnits €650.00
Human cytosolic 5'-nucleotidase II (cN-II)

Provided in stable lyophilized form and shipped without dry ice.
* Pricing updated December 12th, 2015.
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Synonyms: cytosolic 5'-nucleotidase/phosphotransferase, High Km 5’-nucleotidase (hkm-NT), cytosolic purine 5’-nucleotidase (purine 5'-NT), IMP/GMP-specific 5’-nucleotidase

NOVOCIB's human cytosolic IMP/GMP specific 5'-nucleotidase/phosphotransferase II (cN-II) is a pure and active protein of 65kDa cloned by RT-PCR amplification of mRNA extracted from human hepatoma cells and expressed in E.coli. The sequence of the cloned NT5C2 gene (GenBank accession number P49902) was confirmed by DNA sequencing (100% identity).
Cytosolic 5'-nucleotidase II is one of the seven known mammalian nucleotidases(1) that specifically catalyzes the dephosphorylation of 6-hydroxypurine nucleoside 5’-monophosphates (IMP, dIMP, GMP, dGMP) and regulates cellular pool of IMP and GMP(2,3). The enzyme also acts as a phosphotransferase catalyzing the transfer of a phosphate from nucleoside monophosphate to a nucleoside acceptor – preferentially inosine and deoxyinosine. Unlike the other 5’-nucleotidases, cN-II is allosterically regulated by adenine/guanine nucleotides and 2,3-biphosphoglycerate(4).
In addition, cytosolic 5'-nucleotidase II phosphorylates anti-viral and anti-tumour nucleoside analogues such as 2’3’-dideoxyinosine, carbovir(5), acyclovir(6) and ribavirin(7).

Assay condition: Phosphotransferase activity of human recombinant cN-II was followed at 37°C, in 100mM Tris-HCl pH 9; 100mM KCl, 20mM MgCl2, 0.5mM KH2PO4, 5mM DTT, 5mM NAD, 50mU/mL IMPDH (NovoCIB). 5mM GMP was used as phosphate donor and 5mM inosine as - phosphate acceptor. The formation of IMP from inosine was followed at 340nm through coupling to recombinant IMP-dehydrogenase (IMPDH, NovoCIB). In the presence of NAD, IMPDH catalyzes irreversible conversion of IMP to XMP with concomitant reduction of NAD to NADH2 (specific absorbance at 340nm).

Assay condition: 5’-nucleotidase activity of cN-II is followed in an irreversible spectrophotometric assay using coupled purine nucleoside phosphorylase - xanthine dehydrogenase system (2,5 mU/ml each). Assays are carried out at 37°C, at 50mM Tris-HCl pH7,6; 100mM KCl, 20mM MgCl2, 500΅M KH2PO4, 5mM DTT, 119 nM cN-II and various concentrations of IMP. Reaction is followed at 295 nm. IMP is purchased from MP Biochemicals. PNP and XDH are produced and purified by NovoCIB.

References (with links to PubMed)
1. V. Bianchi and J. Spychala (2003): Mammalian 5'-nucleotidases J. Biol. Chem. 278(47), 46195-8
2. S. Allegrini et al. (1997): Bovine cytosolic IMP/GMP-specific 5'-nucleotidase: cloning and expression of active enzyme in Escherichia coli Biochem. J. 328(Pt2), 483-487
3. P. L. Ipata and M. G. Tozzi (2006): Recent advances in structure and function of cytosolic IMP-GMP specific 5'-nucleotidase II (cN-II) Purinergic Signal. 2(4), 669-675
4. J. Spychala et al. (1988): High Km Soluble 5’-nucleotidase from human placenta J. Biol. Chem. 263(35):18759-65
5. M. A. Johnson and A. Fridland (1989): Phosphorylation of 2',3'-dideoxyinosine by cytosolic 5'-nucleotidase of human lymphoid cells Mol. Pharmacol. 36(2), 291-295
6. P. M. Keller (1985): Cytoplasmic 5'-nucleotidase catalyzes acyclovir phosphorylation J. Biol. Chem. 260(15), 8664-7
7. J. Z. Wu et al. (2005): Phosphorylation of ribavirin and viramidine by adenosine kinase and cytosolic 5'-nucleotidase II: Implications for ribavirin metabolism in erythrocytes Antimicrob. Agents Chemother. 49(6): 2164-71

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