Establishment and optimization of a method for screening HIV-1 integrase 3′-processing inhibitor
10.3969/j.issn.1001-1978.2014.10.029
- VernacularTitle:HIV-1整合酶3′端加工抑制剂筛选方法的建立与优化
- Author:
Cuilin LU
;
Xuan ZHANG
;
Jinbiao ZHAN
;
Liumeng YANG
;
Yongtang ZHENG
- Publication Type:Journal Article
- Keywords:
HIV-1;
integrase;
3′-processing;
FRET;
influence factors;
optimization;
inhibitor
- From:
Chinese Pharmacological Bulletin
2014;(10):1469-1473
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish and optimize a method for screening HIV-1 integrase 3′-processing inhibitor. Methods Fluorescence resonance energy transfer ( FRET) was used to create an assay for screening in-tegrase 3′-processing inhibitors; wavelength was de-fined by DNaseⅠ; factors affecting IN activity were optimized, including buffer composition, substrate con-centration, enzyme concentration, metal ion concentra-tion. Results Integrase 3′-processing optimizing reac-tion conditions were buffer 1 , 500 nmol · L-1 sub-strate, 1 μmol·L-1 integrase, 20mmol·L-1 magne-sium ion. Positive drug raltegravir and myricetin could effectively inhibit integrase 3′-processing activity using this assay. Two integrase 3′-processing inhibitors were screened by this method. Conclusion The method for screening HIV-1 integrase 3′-processing inhibitor is successfully established and optimized.
