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标。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-Poll多蛋白至少10个独特的位置，对于病毒粒子的成熟和病毒的传播是必不可少的。因此，它一直是药物设计研究的主要目标。不幸的是，目前药物的医疗效果被证明是短暂的，因为艾滋病毒-1蛋白酶的活性突变变种会产生抗药性。耐药性是识别事件之间的一种微妙变化，即酶与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微妙变化。由于HIV-1蛋白酶在同一活性部位结合底物和抑制物，改变抑制物结合的变化也会改变底物结合。我们先前开发了一个结构原理，解释HIV蛋白酶如何识别其底物，以及耐药突变如何发生在HIV蛋白酶的活性部位，同时仍然保持底物识别。HIV蛋白酶识别底物采用的保守的不对称形状，即“底物包膜”。这使我们意识到，HIV蛋白酶中大多数活性部位的耐药突变发生在抑制物超出共识底物包膜并接触蛋白酶的地方。这些蛋白酶残基是产生耐药性的主要部位，因为它们对抑制剂结合比底物结合更重要。在这项建议中，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及其对蛋白酶抑制剂耐药性的影响，并探讨了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标。不幸的是，目前药物的医疗效果被证明是短暂的，因为艾滋病毒-1蛋白酶的活性突变变种会产生抗药性。耐药性是识别事件之间的一种微妙变化，即HIV蛋白酶对其药物的相对亲和力与其履行其生物学功能的能力之间的微妙变化。在这项建议中，我们研究了这种相互作用，并阐明了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.