A continuous enzyme-coupled assay for triphosphohydrolase activity of HIV-1 restriction factor SAMHD1More about Open Access at the Crick
Authors listLaurence H Arnold Simone Kunzelmann Martin R Webb Ian Taylor
The development of deoxynucleoside triphosphate (dNTP)-based drugs requires a quantitative understanding of any inhibition, activation, or hydrolysis by off-target cellular enzymes. SAMHD1 is a regulatory dNTP-triphosphohydrolase that inhibits HIV-1 replication in human myeloid cells. We describe here an enzyme-coupled assay for quantifying the activation, inhibition, and hydrolysis of dNTPs, nucleotide analogues, and nucleotide analogue inhibitors by triphosphohydrolase enzymes. The assay facilitates mechanistic studies of triphosphohydrolase enzymes and the quantification of off-target effects of nucleotide-based antiviral and chemotherapeutic agents.
Issue number 1
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Publisher website (DOI) 10.1128/aac.03903-14
Europe PubMed Central 25331707
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