A cell-penetrating helical peptide as a potential HIV-1 inhibitor
A cell-penetrating helical peptide as a potential HIV-1 inhibitor
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DOI：
10.1016/j.jmb.2008.02.066
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发表时间：
2008-05-02
影响因子：
5.6
通讯作者：
Ebnath, Asim K.
中科院分区：
文献类型：
--
作者：
Zhang, Hongtao;Zhao, Qian;Ebnath, Asim K.
人类免疫缺陷病毒1型（HIV-1）Gag多聚蛋白的衣壳结构域是病毒组装的关键决定因素，因此是开发艾滋病治疗药物的潜在靶点。最近，一个12聚体的α-螺旋肽（CAI）的报道，破坏不成熟和成熟样衣壳颗粒组装在体外，然而，它未能抑制HIV-1在细胞培养中，由于其无法穿透细胞。同一个小组报告了CAI与衣壳C-末端结构域（C-CA）复合的X射线晶体结构，分辨率为1.7埃。利用这些结构信息，我们利用基于结构的合理设计方法来稳定CAI的α-螺旋结构，并将其转化为细胞穿透肽（CPP）。修饰的肽（NYAD-1）显示增强的α-螺旋性。激光扫描共聚焦显微镜的实验表明，NYAD-1渗透细胞和共定位的Gag多蛋白在其运输到质膜，在那里发生病毒组装。NYAD-1在无细胞和基于细胞的体外系统中破坏了未成熟和成熟样病毒颗粒的组装。NMR化学位移扰动分析将NYAD-1的结合位点映射到HIV-1衣壳的C-末端结构域的残基169 - 191，所述C-末端结构域包含疏水腔和关键二聚化结构域，其结合亲和力比CAI提高。此外，实验数据表明NYAD-1最有可能在进入后阶段靶向衣壳。最重要的是，NYAD-1在低微摩尔效力下抑制细胞培养物中的一大组HIV-1分离株。我们的研究展示了如何使用基于结构的合理设计策略将细胞不可渗透的肽转化为细胞可渗透的肽，该肽在基于细胞的测定中显示活性，而不影响其作用机制。这种概念验证的细胞穿透肽可能有助于验证衣壳作为抗HIV-1药物靶标，并可能有助于设计2008年2月28日接受的肽模拟物和靶向这种蛋白质的小分子药物。(C)2008爱思唯尔有限公司保留所有权利。
The capsid domain of the human immunodeficiency virus type 1 (HIV-1) Gag polyprotein is a critical determinant of virus assembly, and is therefore a potential target for developing drugs for AIDS therapy. Recently, a 12-mer alpha-helical peptide (CAI) was reported to disrupt immature- and mature-like capsid particle assembly in vitro; however, it failed to inhibit HIV-1 in cell culture due to its inability to penetrate cells. The same group reported the X-ray crystal structure of CAI in complex with the C-terminal domain of capsid (C-CA) at a resolution of 1.7 angstrom Using this structural information, we have utilized a structure-based rational design approach to stabilize the alpha-helical structure of CAI and convert it to a cell-penetrating peptide (CPP). The modified peptide (NYAD-1) showed enhanced alpha-helicity. Experiments with laser scanning confocal microscopy indicated that NYAD-1 penetrated cells and colocalized with the Gag polyprotein during its trafficking to the plasma membrane where virus assembly takes place. NYAD-1 disrupted the assembly of both immature- and mature-like virus particles in cell-free and cell-based in vitro systems. NMR chemical shift perturbation analysis mapped the binding site of NYAD-1 to residues 169-191 of the C-terminal domain of HIV-1 capsid encompassing the hydrophobic cavity and the critical dimerization domain with an improved binding affinity over CAI. Furthermore, experimental data indicate that NYAD-1 most likely targets capsid at a post-entry stage. Most significantly, NYAD-1 inhibited a large panel of HIV-1 isolates in cell culture at low micromolar potency. Our study demonstrates how a structure-based rational design strategy can be used to convert a cell-impermeable peptide to a cell-permeable peptide that displays activity in cell-based assays without compromising its mechanism of action. This proof-of-concept cell-penetrating peptide may aid validation of capsid as an anti-HIV-1 drug target and may help in designing peptidomimetics accepted 28 February 2008 and small molecule drugs targeted to this protein. (C) 2008 Elsevier Ltd. All rights reserved.