Antiviral agent based on the non-structural protein targeting the maturation process of HIV-1: expression and susceptibility of chimeric Vpr as a substrate for cleavage by HIV-1 protease.
Antiviral agent based on the non-structural protein targeting the maturation process of HIV-1: expression and susceptibility of chimeric Vpr as a substrate for cleavage by HIV-1 protease.
复制标题
基于针对 HIV-1 成熟过程的非结构蛋白的抗病毒剂：嵌合 Vpr 作为 HIV-1 蛋白酶裂解底物的表达和敏感性。
DOI：
10.1093/protein/13.6.431
复制
发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Srinivasan,A
中科院分区：
文献类型：
--
作者：
Serio,D;Singh,SP;Cartas,MA;Weber,IT;Harrison,RW;Louis,JM;Srinivasan,A
The processing of precursor proteins (Gag and Gag-pol) by the viral protease is absolutely required in order to generate infectious particles. This prompted us to consider novel strategies that target viral maturation. Towards this end, we have engineered an HIV-1 virion associated protein, Vpr, to contain protease cleavage signal sequences from Gag and Gag-pol precursor proteins. We previously reported that virus particles derived from HIV-1 proviral DNA, encoding chimeric Vpr, showed a lack of infectivity, depending on the fusion partner. As an extension of that work, the potential of chimeric Vpr as a substrate for HIV-1 protease was tested utilizing an epitope-based assay. Chimeric Vpr molecules were modified such that the Flag epitope is removed following cleavage, thus allowing us to determine the efficiency of protease cleavage. Following incubation with the protease, the resultant products were analyzed by radioimmunoprecipitation using antibodies directed against the Flag epitope. Densitometric analysis of the autoradiograms showed processing to be both rapid and specific. Further, the analysis of virus particles containing chimeric Vpr by immunoblot showed reactivities to antibodies against the Flag epitope similar to the data observedin vitro. These results suggest that the pseudosubstrate approach may provide another avenue for developing antiviral agents.
