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Antimicrob Agents Chemother. 2015 Jan;59(1):186-92. doi: 10.1128/AAC.03903-14. Epub 2014 Oct 20.

A continuous enzyme-coupled assay for triphosphohydrolase activity of HIV-1 restriction factor SAMHD1.

Author information

1
Division of Molecular Structure, MRC National Institute for Medical Research, London, United Kingdom.
2
Division of Physical Biochemistry, MRC National Institute for Medical Research, London, United Kingdom.
3
Division of Molecular Structure, MRC National Institute for Medical Research, London, United Kingdom itaylor@nimr.mrc.ac.uk.

Abstract

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.

PMID:
25331707
PMCID:
PMC4291348
DOI:
10.1128/AAC.03903-14
[Indexed for MEDLINE]
Free PMC Article

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