HIV Protease Activation and Viral Replication

HIV 蛋白酶激活和病毒复制

• 批准号: 6699055
• 负责人: Andrew H Kaplan
• 金额: $ 27.86万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6699055
• 关键词: HIV infections; antiAIDS agent; aspartic endopeptidases; dimer; enzyme activity; enzyme structure; gag protein; human immunodeficiency virus 1; intermolecular interaction; mutant; protease inhibitor; virus protein; virus replication

DESCRIPTION (provided by applicant): Activation of the HIV-1 protease is an essential step in viral replication. As is the case for all retroviral proteases, enzyme activity requires the formation of protease homodimers. Mutations that block dimerization interfere with the protease function; viral variants encoding non-functional enzymes are aberrantly assembled and non-infectious. In vitro studies of compounds that inhibit the enzyme from dimerizing produce similar results. Despite the invariant nature of protease dimerization across all retroviral systems, important biological and structural questions remain unanswered. First, although structural studies have identified the residues involved in the dimer interface, the role of specific amino acids in promoting and maintaining dimer formation has not been examined. Second, a wealth of structural and biological information suggests that there is a close association between ordered, protease-mediated precursor processing, particle assembly and infectivity. However, the precise effect of protease dimerization and subsequent enzyme activation on particle assembly and infectivity is unclear. Finally, although clinically available substrate-based inhibitors of the HIV protease have made a dramatic impact on disease progression, resistant variants frequently arise in patients treated with these active site-directed compounds. Novel approaches to inhibitor design are urgently needed to develop additional effective therapeutic agents. In the studies described below, we will define the interactions critical for maintenance of the HIV protease dimer and the role that the final structure plays in viral replication. Further, we will explore novel strategies for inhibition of the viral protease through disrupting dimer formation. Overall, our studies will provide important insights into protease structure, enzyme function and retroviral biology. Specifically, we will: I. Define the role of individual amino acids in dimer formation. II. Characterize the effect of substitutions in the F-IIV protease dimer interface on viral replication. IlI. Characterize HIV protease activation within GagPol as a potential target for inhibition.

描述（由申请人提供）：HIV-1蛋白酶的激活是病毒复制的重要步骤。与所有逆转录病毒蛋白酶一样，酶活性需要形成蛋白酶同二聚体。阻断二聚化干扰蛋白酶功能的突变；编码非功能性酶的病毒变体被异常组装且不感染。对抑制酶二聚体的化合物的体外研究产生相似的结果。尽管在所有逆转录病毒系统中蛋白酶二聚化的不变性质，但重要的生物学和结构性问题仍未得到解决。首先，尽管结构研究已经确定了二聚体界面中涉及的残基，但尚未研究特定氨基酸在促进和维持二聚体形成中的作用。其次，大量的结构和生物学信息表明，有序，蛋白酶介导的前体加工，颗粒组装和感染性之间存在密切的关联。然而，蛋白酶二聚化和随后的酶激活对颗粒组装和感染性的确切作用尚不清楚。最后，尽管临床上可用的基于底物的HIV蛋白酶抑制剂对疾病进展产生了巨大影响，但在接受这些活性部位定向化合物治疗的患者中，经常出现抗性变异。迫切需要采用抑制剂设计的新型方法来开发其他有效的治疗剂。在下面描述的研究中，我们将定义对维持HIV蛋白酶二聚体至关重要的相互作用以及最终结构在病毒复制中所扮演的作用。此外，我们将通过破坏二聚体的形成来探索抑制病毒蛋白酶的新型策略。总体而言，我们的研究将为蛋白酶结构，酶功能和逆转录病毒生物学提供重要的见解。具体来说，我们将： I.定义单个氨基酸在二聚体形成中的作用。 ii。表征F-IIV蛋白酶二聚体界面中取代对病毒复制的影响。 ili。将Gagpol中的HIV蛋白酶激活表征为抑制的潜在靶标。