Human cytosolic 5'-nucleotidase II (cN-II) (E.C.3.1.3.5)

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Pricing updated on July 23, 2024.

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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 specifically catalyzes the dephosphorylation of 6-hydroxypurine nucleoside 5'-monophosphates (IMP, dIMP, GMP, dGMP) and regulates cellular pools of IMP and GMP. The enzyme also acts as a phosphotransferase catalyzing phosphate transfer from nucleoside monophosphates to nucleoside acceptors, preferentially inosine and deoxyinosine. Unlike other 5'-nucleotidases, cN-II is allosterically regulated by adenine/guanine nucleotides and 2,3-biphosphoglycerate.
In addition, cN-II phosphorylates antiviral and antitumor nucleoside analogues such as 2'3'-dideoxyinosine, carbovir, acyclovir, and ribavirin.

Synonyms: cytosolic 5'-nucleotidase/phosphotransferase, High Km 5'-nucleotidase (HKM-NT), cytosolic purine 5'-nucleotidase (purine 5'-NT), IMP/GMP-specific 5'-nucleotidase.

Download Human cytosolic 5'-nucleotidase II (User manual)

Assay condition:

  • Temperature: 37°C
  • Buffer: 100mM Tris-HCl pH 9
  • Reagents: 100mM KCl, 20mM MgCl2, 0.5mM KH2PO4, 5mM DTT, etc.
  • Phosphate donor: 5mM GMP
  • Phosphate acceptor: 5mM inosine
  • Method: Coupling to recombinant IMP-dehydrogenase (IMPDH, NovoCIB)
Illustration of phosphotransferase activity of cN-II enzyme

Assay condition:

  • Temperature: 37°C
  • Buffer: 50mM Tris-HCl pH7.6
  • Reagents: 100mM KCl, 20mM MgCl2, 500µM KH2PO4, etc.
  • Method: Coupling with purine nucleoside phosphorylase - xanthine dehydrogenase system.
Illustration of cN-II enzyme kinetics
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.