Development of a high-throughput assay for the HIV-1 integrase disintegration reaction
He, H.; Liu, B.; Zhang, X.; Chen, W.; Wang, C.
Science China. Life Sciences 53(2): 241-247
2010
ISSN/ISBN: 1869-1889 PMID: 20596834 DOI: 10.1007/s11427-010-0006-7Document Number: 212811
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.
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