Disruption of the HIV-1 protease dimer with interface peptides:: Structural studies using NMR spectroscopy combined with [2-13C]-Trp selective labeling
Disruption of the HIV-1 protease dimer with interface peptides:: Structural studies using NMR spectroscopy combined with [2-13C]-Trp selective labeling
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DOI：
10.1002/bip.20685
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发表时间：
2007-01-01
期刊：
影响因子：
2.9
通讯作者：
Giralt, Ernest
中科院分区：
文献类型：
--
作者：
Frutos, Silvia;Rodriguez-Mias, Ricard A.;Giralt, Ernest
HIV-1蛋白酶（HIV-1 PR）由逆转录病毒编码，是加工gag和pol多聚蛋白前体所必需的，因此它对感染性病毒颗粒的产生至关重要。在体外抑制酶的结果在后代病毒粒子的生产是不成熟的和非感染性的，这表明其作为艾滋病的治疗靶点的潜力。虽然现在有许多有效的蛋白酶抑制剂药物，但由于HIV-1 PR突变而对这些药物产生耐药性，迫切需要新的HIV-1 PR抑制方法。虽然酶通常通过阻断活性位点而失活，但二聚体HIV-1 PR的结构允许替代的抑制机制。由于活性位点由两个半酶形成，它们通过涉及两个单体的N-和C-末端的四链反平行β-折叠连接，因此酶活性可以通过靶向相对不含突变的区域中的二聚体界面的试剂消除，这些突变会干扰功能性HIV-1 PR二聚体的形成或稳定性。几个小组已经探索了这种策略，他们靶向四链反平行β折叠，贡献了近75%的二聚化能量。主要基于动力学分析，证明了对应于其几种模拟物的天然单体N-或C-末端的界面肽作为二聚化抑制剂。然而，据我们所知，迄今为止还没有对酶抑制剂复合物进行X射线晶体学和NMR结构研究。在这篇文章中，我们报告了使用选择性Trp侧链标记通过NMR对HIV-1 PR的二聚化抑制的结构研究。© 2007 Wiley Periodicals，Inc. Biopolymers（Pept Sci）88：164-173，2007。这篇文章最初作为公认的预印本在线发表。“在线发布”日期对应于预印本版本。您可以通过向Biopolymers编辑部发送电子邮件（biopolymers@wiley.com）索取预印本的副本
HIV‐1 protease (HIV‐1 PR), which is encoded by retroviruses, is required for the processing of gag and pol polyprotein precursors, hence it is essential for the production of infectious viral particles. In vitro inhibition of the enzyme results in the production of progeny virions that are immature and noninfectious, suggesting its potential as a therapeutic target for AIDS. Although a number of potent protease inhibitor drugs are now available, the onset of resistance to these agents due to mutations in HIV‐1 PR has created an urgent need for new means of HIV‐1 PR inhibition. Whereas enzymes are usually inactivated by blocking of the active site, the structure of dimeric HIV‐1 PR allows an alternative inhibitory mechanism. Since the active site is formed by two half‐enzymes, which are connected by a four‐stranded antiparallel β‐sheet involving the N‐ and C‐ termini of both monomers, enzyme activity can be abolished by reagents targeting the dimer interface in a region relatively free of mutations would interfere with formation or stability of the functional HIV‐1 PR dimer. This strategy has been explored by several groups who targeted the four‐stranded antiparallel β‐sheet that contributes close to 75% of the dimerization energy. Interface peptides corresponding to native monomer N‐ or C‐termini of several of their mimetics demonstrated, mainly on the basis of kinetic analyses, to act as dimerization inhibitors. However, to the best of our knowledge, neither X‐ray crystallography nor NMR structural studies of the enzyme‐inhibitor complex have been performed to date. In this article we report a structural study of the dimerization inhibition of HIV‐1 PR by NMR using selective Trp side chain labeling. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 88: 164–173, 2007.This article was originally published online as an accepted preprint. The ‘Published Online’ date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com