MUTATIONS EFFECTS ON INHIBITION OF HIV PROTEASE

描述(改编自摘要)：本项目的目标是 了解突变对基因表达的催化和抑制的影响 艾滋病毒蛋白水解酶。一大批有效的抗HIV药物 目前，许多实验室已经合成并测试了蛋白酶。一个 目前，这些药物中几乎没有处于临床试验中。然而，越来越多的人， 最近的证据支持这样一种观点，即艾滋病毒对 通过其蛋白酶的突变而产生的蛋白酶抑制物。有能力 分别调节蛋白水解酶活性和抑制率 突变的敏感性是抗药性的基础。因此， 目前项目中的研究人员正在寻求突变如何产生的基本知识 HIV蛋白酶在催化和抑制方面的变化是如何 变化会转化为抵抗。他们的项目有两个 组成部分：(1)了解活性和抑制作用是如何变化的 由于HIV蛋白酶的突变；以及(2)确定晶体 突变的HIV蛋白水解酶抑制剂复合体的结构。突变酶 具有“抗抑制”特性的是那些保留了 催化效率与野生型酶相当，但 对抑制剂的敏感度显著降低。在之前的研究中，这些 研究人员在几个突变体中观察到了这些特征。AS 本项目的第一部分，首席调查员提出了一个 广泛研究潜在的耐药突变位点，多个 突变和“邻近”突变。许多分析工具将是 酶和抑制动力学，以及可预测的动力学模型 在抑制剂存在的情况下处理突变酶的活性。 预测缓蚀剂相对易损性的动力学工具 对抗药性将进行进一步测试。认识到结构性的 信息对于解释两者之间的关系至关重要 突变和抗药性动力学行为导致的结构变化 调查人员提出作为该项目的第二个组成部分 最有趣的突变蛋白水解酶-抑制物对的结晶 用X-射线单晶衍射法测定了其晶体结构。 抗药性突变酶的新结构将被比较。 仔细研究野生型酶和非抗性突变体的结构 酵素。这些比较将为动力学数据提供洞察力 在组件#1中生成。

DESCRIPTION (adapted from the Abstract): The goal of this project is to understand the effects of mutations on the catalysis and inhibition of the HIV protease. A large number of potent inhibitors against HIV protease have now been synthesized and tested by many laboratories. A few of these are currently in clinical trials. However, increasingly, recent evidence supports the view that HIV acquires resistance to protease inhibitors by mutation of its protease. The ability to modulate separately the proteolytic activity and the inhibition sensitivity by mutation is the basis of resistance. Thus, the researchers in the current project seek basic knowledge of how mutations of HIV protease render changes in catalysis and inhibition how these changes are translated into resistance. Their project has two components: (1) to understand how the activity and inhibition are changed due to mutations of HIV protease; and (2) to determine the crystal structures of mutant HIV protease- inhibitor complexes. Mutant enzymes with characteristics of "inhibition resistance" are those which retain catalytic efficiency comparable to the wild-type enzyme but are significantly less sensitive to inhibitors. In previous studies, these researchers have observed these characteristics in several mutants. As Component #1 of this project, the Principal Investigator proposes an extensive study of potential resistant mutation sites, multiple mutations, and "neighboring" mutations. The many analytic tool will be enzyme and inhibition kinetics and a kinetic model which can predict the processing activities of mutant enzymes in the presence of inhibitors. Kinetic tools for predicting the relative vulnerability of inhibitors to resistance will be tested further. Recognizing that structural information is essential for interpreting the relationship between the structural changes by mutations and resistant kinetic behavior, the Investigator proposes as Component #2 of this project the crystallization of the most interesting mutant protease-inhibitor pairs and the determination of the crystal structures by X-ray crystallography. The new structures for the resistant mutant enzymes will be compared carefully to the structures of wild-type enzyme and non-resistant mutant enzymes. These comparison will provide insights for the kinetic data generation in Component #1.