一种用于检测HIV-1限制因子SAMHD1三磷酸水解酶活性的连续酶联测定法。
A continuous enzyme-coupled assay for triphosphohydrolase activity of HIV-1 restriction factor SAMHD1.
作者信息
Arnold Laurence H, Kunzelmann Simone, Webb Martin R, Taylor Ian A
机构信息
Division of Molecular Structure, MRC National Institute for Medical Research, London, United Kingdom.
Division of Physical Biochemistry, MRC National Institute for Medical Research, London, United Kingdom.
出版信息
Antimicrob Agents Chemother. 2015 Jan;59(1):186-92. doi: 10.1128/AAC.03903-14. Epub 2014 Oct 20.
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.
基于脱氧核苷三磷酸（dNTP）的药物研发需要对脱靶细胞酶的任何抑制、激活或水解作用有定量的认识。SAMHD1是一种调节性dNTP三磷酸水解酶，可抑制HIV-1在人髓细胞中的复制。我们在此描述一种酶联测定法，用于定量三磷酸水解酶对dNTP、核苷酸类似物和核苷酸类似物抑制剂的激活、抑制和水解作用。该测定法有助于对三磷酸水解酶进行机制研究，并对基于核苷酸的抗病毒和化疗药物的脱靶效应进行定量分析。
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