Development of a high-throughput assay for the HIV-1 integrase disintegration reaction.
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.
He
HongQiu
H
College of Life Science and Bioengineering, Beijing University of Technology, Beijing, 100124, China.
Liu
Bin
B
Zhang
XiaoYi
X
Chen
WeiZu
W
Wang
CunXin
C
D002384:Catalysis; D020134:Catalytic Domain; D002413:Cations, Divalent; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019427:HIV Integrase / Q000378:metabolism*; D006801:Humans; D019429:Integrase Inhibitors / Q000032:analysis
D002413:Cations, Divalent; D019429:Integrase Inhibitors; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1