Human Immunodeficiency Virus Proteinase

人类免疫缺陷病毒蛋白酶

• 批准号: 6510426
• 负责人: Ben M. Dunn
• 金额: $ 27.22万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6510426
• 关键词: antiAIDS agent; chimeric proteins; drug resistance; endopeptidases; enzyme activity; gag protein; gene mutation; genetic polymorphism; human immunodeficiency virus 1; human tissue; microorganism culture; nucleotidyltransferase; protease inhibitor; recombinant virus; virus replication

DESCRIPTION (Provided by applicant): Antiretroviral therapy with a combination of RT and PR inhibitors has had a profound effect upon the management of HIV-1 infection. However, under drug pressure, the virus is able to evolve into forms that are resistant to the available drug regimens. Thus, drug resistance has become the most significant challenge to AIDS therapy. We have been studying a pediatric population undergoing a clinical trial with protease inhibitors in combination with RT inhibitors. We have discovered that evolution of the protease sequence is accompanied by evolution of the sequence of the cleavage sites within Gag/Pol, thus affecting the efficiency of processing. Furthermore, the evolution of protease is accompanied by alterations in cleavage specificity. We hypothesize that the Gag/Pol cleavage sites and PR form a functional unit in which sequence evolution may be co- dependent. In our renewal period, we will exploit these discoveries to gain additional understanding of the mechanisms of Gag/Pol processing. To this end, we will pursue the following specific aims: Specific Aim I will analyze natural Gag/Pol alleles for processing phenotype in a bacterial expression developed in the current period of support. We will also map the determinants by preparation of chimeric gag/pol constructs and analyze replication of recombinant virus with natural or chimeric gag/pol regions. Specific Aim 2 will study the growth of recombinant virus in the presence of anti-protease drugs in culture. In addition, new protease alleles will be subcloned and expressed for analysis by studies of substrate specificity using a combinatorial library of substrates, and by analysis of the binding of inhibitors. Specific Aim 3 will study the effects of changes in the sequence of Gag processing sites on the efficiency of processing. We will prepare peptides representing products of Gag/Pol processing and test their ability to inhibit the enzyme. We will also conduct structural analyses of fusions of Gag/Pol proteins with an inactive form of HIV PR in order to determine the role of structural organization in the processing events.

描述（由申请人提供）：抗逆转录病毒治疗与联合 RT和PR抑制剂对HIV-1的管理产生了深远的影响 感染然而，在药物压力下，病毒能够进化成 对现有的药物治疗方案有抵抗力。因此，耐药性 成为艾滋病治疗的最大挑战。我们一直在研究 正在接受蛋白酶抑制剂临床试验的儿科人群， 与RT抑制剂的组合。我们发现， 蛋白酶序列伴随着切割序列的进化 在Gag/Pol内的站点，从而影响处理效率。此外，委员会认为， 蛋白酶的进化伴随着切割的改变 的特异性我们假设Gag/Pol切割位点和PR形成了一个 序列进化可能相互依赖的功能单位。在我们 在更新期内，我们将利用这些发现， 了解Gag/Pol加工的机制。为此我们将 追求以下具体目标：具体目标我将分析自然加格/波尔 在细菌表达中开发的加工表型的等位基因， 目前的支持。我们还将绘制决定因素， 嵌合gag/pol构建体并分析重组病毒复制 天然或嵌合的gag/pol区。具体目标2将研究的增长 重组病毒在培养物中存在抗蛋白酶药物的情况下。在 此外，新的蛋白酶等位基因将被亚克隆和表达，用于分析， 使用底物组合文库的底物特异性研究， 并通过分析抑制剂的结合。具体目标3将研究 的影响，在加格处理网站的顺序变化的效率， 处理.我们将制备代表Gag/Pol产物的肽 加工并测试它们抑制酶的能力。我们还将进行 Gag/Pol蛋白与HIV非活性形式融合的结构分析 以确定PR结构组织在处理中的作用 事件