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(E.C.3.1.3.5) * Pricing updated September 23rd, 2011. Prices are subject to change without notice. Shipping charges are not included. ** One unit of 5’nucleotidase converts 1.0 µmole of IMP to inosine per minute at pH 7.6 at 37°C, as measured by a coupled PNP/XDH enzyme system. Specific Activity: ≥ 150 mU/mg protein. For other size, please contact us Click to order or to get further information |
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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 (IMP/GMP-specific 5'-NT)
NOVOCIB's human cytosolic IMP/GMP specific 5'-nucleotidase/phosphotransferase II (cN-II) is a recombinant protein of ca. 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).
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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).
 Unit Definition: One unit of 5’nucleotidase converts 1.0 µmole of IMP to inosine per minute at pH 7.6 at 37°C, as measured by a coupled PNP/XDH enzyme system in the presence of 20mM MgCl2, 5mM DTT, 500µM KH2PO4, and 1,25mM IMP. Specific Activity: ≥ 0.150 unit/mg protein. Purity: controlled by 10% AA SDS-PAGE. |
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, 500µM 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.
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