Elucidating the Interplay of Protease Substrate Recognition and Drug Resistance

描述（申请人提供）：HIV-1蛋白酶是治疗HIV-1感染最有效的抗病毒药物的靶点。所有这些药物都来自成功的基于结构的设计研究。该酶切割病毒gag-pol多聚蛋白的至少10个独特位点，并且对于病毒体的成熟以及因此病毒的传播是必需的。因此，它已成为药物设计研究的主要目标。不幸的是，目前药物的医疗效力被证明是短暂的，因为HIV-1蛋白酶的可行突变变体赋予耐药性。耐药性是酶结合抑制剂的相对亲和力与其结合和切割底物的能力之间的识别事件平衡的微妙变化。由于HIV-1蛋白酶在相同的活性位点结合底物和抑制剂，改变抑制剂结合的变化也改变底物结合。我们以前开发了一种结构原理，解释了HIV蛋白酶如何识别其底物，以及HIV蛋白酶活性位点内如何发生耐药突变，同时仍保持底物识别。HIV蛋白酶识别底物所采用的保守的不对称形状，即“底物包膜”。这使我们认识到，HIV蛋白酶内的大多数活性位点耐药突变发生在抑制剂突出超过共有底物包膜并接触蛋白酶的地方。这些蛋白酶残基是耐药性发生的主要位置，因为它们对抑制剂结合比对底物结合更重要。在这个提议中，我们阐明了药物诱导的HIV-1蛋白酶和Gag内的共同进化的相互依赖性及其对蛋白酶抑制剂耐药性的影响，并研究如何将底物包膜扩展到其他系统。 公共卫生相关性：HIV-1蛋白酶是治疗HIV-1感染最有效的抗病毒药物的靶点。不幸的是，目前药物的医疗效力被证明是短暂的，因为HIV-1蛋白酶的可行突变变体赋予耐药性。耐药性是HIV蛋白酶对其药物的相对亲和力与其执行生物学功能的能力之间识别事件平衡的微妙变化。在这项提案中，我们调查这种相互作用，并阐明对不同药物的耐药性的影响。

DESCRIPTION (provided by applicant): HIV-1 protease is the target of the most effective anti-viral drugs for the treatment of HIV-1 infection. All these drugs derive from successful structure-based design studies. The enzyme cleaves the viral gag-pol polyprotein at least ten unique sites and is essential for maturation of the virion and thus the spread of the virus. Therefore, it has been a prime target for drug design research. Unfortunately the medical efficacy of the current drugs is proving to be short lived, as viable mutant variants of HIV-1 protease confer drug resistance. Drug resistance is a subtle change in the balance of recognition events, between the relative affinity of the enzyme to bind inhibitors and its ability to bind and cleave substrates. Since HIV-1 protease binds substrates and inhibitors at the same active site, a change that alters inhibitor binding also alters substrate binding. We previously developed a structural rationale that explains how HIV protease recognizes its substrates and how drug resistant mutations occur within the active site of HIV protease, while still maintaining substrate recognition. HIV protease recognizes a conserved asymmetric shape that the substrates adopt, the "substrate envelope". This led us to the realization that most active-site drug-resistant mutations within HIV protease occur where the inhibitors protrude beyond the consensus substrate envelope and contact the protease. Those protease residues are prime positions for drug resistance to occur, as they are more important for inhibitor binding than for substrate binding. In this proposal we elucidate the interdependence of drug-induced co-evolution of HIV-1 protease and within Gag and its impact on protease inhibitor drug resistance and investigate how to extend the substrate envelope to other systems. PUBLIC HEALTH RELEVANCE: HIV-1 protease is the target of the most effective anti-viral drugs for the treatment of HIV-1 infection. Unfortunately the medical efficacy of the current drugs is proving to be short lived, as viable mutant variants of HIV-1 protease confer drug resistance. Drug resistance is a subtle change in the balance of recognition events, between the relative affinity of HIV protease to its drugs and its ability perform its biological function. In this proposal we investigate this interplay and elucidate the impact on drug resistance to different drugs.