Development of a high-throughput assay for the HIV-1 integrase disintegration reaction

Abstract

Both HIV-1 integrase (IN) and the central catalytic domain of IN (IN-CCD) catalyze the disintegration reaction in vitro. In this study, IN and IN-CCD proteins were expressed and purified, and a high-throughput format enzyme-linked immunosorbent assay (ELISA) was developed for the disintegration reaction. IN exhibited a marked preference for Mn(2+) over Mg(2+) as the divalent cation cofactor in disintegration. Baicalein, a known IN inhibitor, was found to be an IN-CCD inhibitor. The assay is sensitive and specific for the study of disintegration reaction as well as for the in vitro identification of antiviral drugs targeting IN, especially targeting IN-CCD.

Bioinformatics Accelerates Drug Repositioning*

“…发现始于偶然，新药巨额的研发成本则迫使生物制 药产业主动地去寻找旧药的新用途，以使现有的药 物得到最大化的利用，同时也使制药厂商的利润最 大化． 比较普遍的研发思路是：首先构建已获批准 的药物库(librariesofapproveddrugs)，利用高通量 的筛选技术平台 [13] 大规模地筛查潜在的药靶，筛查 到药物潜在的新用途后，再经过科学的验证体系进 行验证，并通过各期临床试验，完成市场前景、知 识产权等多方面的评估，最后重新获得药品监管部 门的批准，进入新的市场．…”

A glimpse of enzymology within the idea of systems

“…High-throughput screening assays with adequate sensitivity, reproducibility, and accuracy have been indispensable for the discovery of drugs targeting HIV-1 integrase. In a recent research article, He and coworkers [26] developed a novel method for measuring the disintegration activity of HIV-1 integrase. This method is rapid, sensitive and, most importantly, readily adapted to a high-throughput format, and hence is likely to be applicable to antiviral drug screening.…”