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Issue 34, 2010
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Rapid determination of adenosine deaminase kinetics using fast-scan cyclic voltammetry

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Abstract

Adenosine deaminase is an enzyme involved in purine metabolism and its inhibitors are used as anticancer and antiviral drugs. In this study, we show that fast-scan cyclic voltammetry at carbon-fiber microelectrodes can be used to study the kinetics of adenosine deaminase by electrochemically monitoring decreases in adenosine concentration. Buffer and salt concentrations were shown to affect the enzyme kinetics and the inhibition by erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and deoxycoformycin (DCF). In a Tris buffer containing salts that mimic cerebrospinal fluid, EHNA and DCF showed non-competitive inhibition with a Ki of 1.7 ± 0.6 nM and 1.2 ± 0.2 nM, respectively. However, removing the divalent cations from the Tris buffer caused the inhibition to be competitive and reduced the Ki for DCF by two orders of magnitude. In phosphate-buffered saline, the Ki was 1.0 ± 0.2 nM for EHNA and 3.6 ± 0.3 pM for DCF, similar to literature values. Adenosine deaminase was also competitively inhibited by AgNO3, showing it is susceptible to silver toxicity. Caffeine was found to increase adenosine deaminase activity. This is a fast, easy method for screening drug effects on enzyme kinetics and could be applied to other enzymatic reactions where there is a significant difference in the electroactivity of the reactant and product.

Graphical abstract: Rapid determination of adenosine deaminase kinetics using fast-scan cyclic voltammetry

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Publication details

The article was received on 20 Apr 2010, accepted on 28 May 2010 and first published on 24 Jun 2010


Article type: Paper
DOI: 10.1039/C0CP00294A
Citation: Phys. Chem. Chem. Phys., 2010,12, 10027-10032
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    Rapid determination of adenosine deaminase kinetics using fast-scan cyclic voltammetry

    Y. Xu and B. J. Venton, Phys. Chem. Chem. Phys., 2010, 12, 10027
    DOI: 10.1039/C0CP00294A

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