Human Adenosine Kinase (ADK, EC 2.7.1.20)

Synonyms: ADK, Adenosine 5'-phosphotransferase

NOVOCIB's human adenosine kinase is an active and purified, 345-aa short form (39kDa) ADK protein cloned by RT-PCR amplification of mRNA extracted from human hepatoma cells and expressed in E.coli. The sequence of the cloned ADK (GenBank accession number U50196) was confirmed by DNA sequencing (100% identity).

Adenosine kinase catalyzes the transfer of γ-phosphate from ATP to the 5'-hydroxyl of adenosine, a key modulator of central nervous system functions and a signal molecule involved in hypoxia, inflammation, and nociception. Together with adenosine deaminase, adenosine kinase regulates intra- and extracellular adenosine concentration.

Inhibition of adenosine kinase selectively increases local adenosine concentrations and reduces seizure susceptibility and nociception in vivo. ADK dysfunction is implicated in diabetes, epilepsy, and cancer. Consequently, ADK is a rational therapeutic target for drug discovery, and adenosine-regulating drugs have been tested as analgesic, anti-inflammatory, and neuroprotective agents.

ADK is also responsible for phosphorylation and clinical activity of several therapeutic nucleosides, including the antiviral drug ribavirin, immunosuppressive drug mizoribine, and anticancer C-nucleoside tiazofurin.

For rapid evaluation of substrate properties of novel nucleoside analogs for human adenosine kinase see our PRECICE® ADK Phosphorylation Assay Kit.

For rapid HTS search of novel ADK inhibitors see our PRECICE® ADK Assay Kit.

Reaction schema of human adenosine kinase

Adenosine Kinase

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Last updated on : November 7th, 2025.

Bulk quantity available

Kit is provided in stable lyophilized form and shipped without dry ice

You can ask us for a quotation here or write at contact@novocib.com

Km and Vmax Data for Nucleoside Analogues

Validated enzymatic parameters for human recombinant adenosine kinase (ADK) compared with published values:

Substrate Km (µM, Novocib) Kcat (min⁻¹, Novocib) Km (µM, Published) Kcat (min⁻¹, Published) Reference
Adenosine 11 1.5 3.2 / 0.150 13 Wu et al. 2005; Yamada et al. 1981
Ribavirin 328 1.9 540 1.8 Wu et al. 2005
Deoxyadenosine 295 3.4 360 - Yamada et al. 1981
Tubercidine 12 2.2 - - -
Inosine 1758 2.6 - - -

Assay Conditions and Enzyme Characteristics

SDS-PAGE gel showing purified human adenosine kinase

Unit Definition: One unit of adenosine kinase converts 1.0 µmole of inosine and ATP to IMP and ADP per minute at pH 8 at 37°C, as measured by a coupled IMPDH enzyme system.

Specific Activity: ≥ 0.200 unit/mg protein.

Purity: controlled by 12% AA SDS-PAGE.

Assay condition: Enzymatic activity of adenosine kinase with particular nucleoside substrate is measured by spectrophotometric assays in a coupled lactate dehydrogenase / pyruvate kinase system. Assays were carried out at 37°C, at 50mM Tris-HCl pH7.6; 50mM KCl, 5mM MgCl2, 2.5mM ATP, 0.1mM NADH, 1mM phosphoenolpyruvate, 1mM DTT, PK 5U/ml, LDH 5U/ml. Reaction was followed in an iEMS Reader MF (Labsystems) microtiter plate reader at 340nm.

Kinetics graph of ribavirin phosphorylation by adenosine kinase

Frequently Asked Questions

ADK phosphorylates adenosine and several nucleoside analogues including ribavirin, tubercidine, mizoribine, and tiazofurin.

Purity is verified by SDS-PAGE analysis and specific activity measurements to ensure consistent quality.

Yes, client-specified alterations and custom nucleoside phosphorylation assays are available upon request.

ADK is a validated target for analgesic, anti-inflammatory, and neuroprotective drugs. It is also essential for the activation of therapeutic nucleosides such as ribavirin, mizoribine, and tiazofurin, making it highly relevant in antiviral, immunosuppressive, and anticancer drug development.
Scientific Works citing NOVOCIB Adenosine Kinase and PRECICE® ADK Assay kits:
  1. Wu JZ, Larson G, Walker H, Shim JH, Hong Z.
    Phosphorylation of ribavirin and viramidine by adenosine kinase and cytosolic 5'-nucleotidase II: Implications for ribavirin metabolism in erythrocytes. Antimicrob Agents Chemother. 2005;49(6):2164-2171.
  2. Yamada Y, Goto H, Ogasawara N.
    Adenosine kinase from human liver. Biochim Biophys Acta. 1981;660(1):36-43.
  3. Orlicka-Płocka M, Fedoruk-Wyszomirska A, Gurda-Wozna D, Pawelczak P, Krawczyk P, Giel-Pietraszuk M, Framski G, Ostrowski T, Wyszko E.
    Implications of Oxidative Stress in Glioblastoma Multiforme Following Treatment with Purine Derivatives. Antioxidants. 2021;10:950.
  4. Johnson LM, Smith OJ, Hahn DA, Baer CF.
    Short-term heritable variation overwhelms 200 generations of mutational variance for metabolic traits in Caenorhabditis elegans. Evolution. 2020;74(11):2451-2464.
  5. Danielyan K, Vardanyan RD, Simonyan A, Sagyan AS.
    The sole role of PRPS-1 in the regenerative processes after experimental stroke. Poster#: 459.07/A7. Society for Neuroscience Annual Meeting. Washington, DC, 2017.
  6. Nayar U, Sadek J, Reichel J, Hernandez-Hopkins D, Akar G, Barelli PJ, Sahai MA, Zhou H, Totonchy J, Jayabalan D, Niesvizky R, Guasparri I, Hassane D, Liu Y, Sei S, Shoemaker RH, Warren JD, Elemento O, Kaye KM, Cesarman E.
    Identification of a nucleoside analog active against adenosine kinase-expressing plasma cell malignancies. J Clin Invest. 2017;127(6):2066-2080.
  7. Toti KS, Osborne D, Ciancetta A, Boison D, Jacobson KA.
    South (S)- and North (N)-Methanocarba-7-Deazaadenosine Analogues as Inhibitors of Human Adenosine Kinase. J Med Chem. 2016;59(14):6860-6877.
  8. Bjursell MK, Blom HJ, Cayuela JA, Engvall ML, Lesko N, Balasubramaniam S, Brandberg G, Halldin M, Falkenberg M, Jakobs C, Smith D, Struys E, von Döbeln U, Gustafsson CM, Lundeberg J, Wedell A.
    Adenosine Kinase Deficiency Disrupts the Methionine Cycle and Causes Hypermethioninemia, Encephalopathy, and Abnormal Liver Function. Am J Hum Genet. 2011;89(4):507-515.
  9. Cesarman E, Nayar U, Warren JD, Sadek J.
    Novel nucleoside analogs and use thereof in therapeutic treatment. Patent application filed by Cornell University. US20190225643A1.