MUTATIONS EFFECTS ON INHIBITION OF HIV PROTEASE

描述（改编自摘要）：本项目的目标是 了解突变对催化和抑制的影响， HIV蛋白酶大量有效的艾滋病毒抑制剂 蛋白酶现在已经被许多实验室合成和测试。一 目前这些药物中只有少数处于临床试验阶段。然而，越来越多的， 最近的证据支持这样的观点，即艾滋病毒获得了对 蛋白酶抑制剂通过突变其蛋白酶。的能力 分别调节蛋白水解活性和抑制 突变引起的敏感性是耐药性的基础。因此 当前项目的研究人员寻求突变如何 HIV蛋白酶的催化和抑制作用发生变化， 变化转化为阻力。他们的项目有两个 成分：（1）了解活性和抑制是如何变化的 由于HIV蛋白酶的突变;和（2）确定晶体 突变的HIV蛋白酶-抑制剂复合物的结构。突变酶 具有“抗抑制性”特征的是那些保留 催化效率与野生型酶相当，但 对抑制剂的敏感性显著降低。在以前的研究中，这些 研究人员已经在几种突变体中观察到这些特征。作为 该项目的第一部分，主要研究者提出了一项 对潜在耐药突变位点的广泛研究， 突变和“邻近”突变。许多分析工具将是 酶和抑制动力学和动力学模型可以预测 在抑制剂存在下突变酶的加工活性。 预测抑制剂相对易损性的动力学工具 抵抗力将进一步测试。认识到，结构 信息对于解释 结构变化的突变和耐药动力学行为， 研究者建议作为本项目的组件#2， 最有趣的突变蛋白酶-抑制剂对的结晶 并通过X射线晶体学测定晶体结构。 将比较抗性突变酶的新结构 仔细观察野生型酶和非抗性突变体的结构 内切酶这些比较将为动力学数据提供见解 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.