Design of a novel peptide inhibitor of HIV fusion that disrupts the internal trimeric coiled-coil of gp41
Design of a novel peptide inhibitor of HIV fusion that disrupts the internal trimeric coiled-coil of gp41
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DOI：
10.1074/jbc.m201453200
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发表时间：
2002-04-19
影响因子：
4.8
通讯作者：
Clore, GM
中科院分区：
文献类型：
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作者：
Bewley, CA;Louis, JM;Clore, GM
来自人类免疫缺陷病毒（HIV）的gp41的前发夹中间体是两类融合抑制剂的靶标，它们结合c端区域或n端螺旋的三聚体卷曲线圈，从而阻止发夹融合三聚体的形成。采用合理的设计，从包含gp41外结构域n端螺旋的亲本N36肽（HIV-1包膜的546-581残基）中衍生出N36（Mut(e,g)）和N36（Mut(a,d)）两个36位残基肽，通过分析性超离心和CD进行表征，并使用基于牛痘病毒的定量融合试验评估它们抑制HIV融合的能力。N36（Mut(e,g)）包含9个氨基酸取代，旨在破坏与gp41 c端区域的相互作用，同时保留控制三聚体卷曲线圈形成的接触。N36（Mut(a,d)）包含9个取代，旨在阻止三聚体卷曲线圈的形成，但保留与gp41的c端区相互作用的残基。N36（Mut(a,d)）是一个单体，大部分是随机线圈，不与gp41 c端区衍生的C34肽（残基628-661）相互作用，也不抑制融合。因此，三聚体线圈结构是gp41 c端相互作用的先决条件。N36（Mut(e,g)）在溶液中形成单分散的螺旋三聚体，不与C34相互作用，但抑制融合的效果比母体N36肽高50倍（IC50 -类似于308 nM，而类似于16 nM）。这些结果表明N36（Mut(e,g)）通过破坏发夹前中间体中n端螺旋的同三聚体线圈形成异三聚体。因此，N36（Mut(e,g)）代表了一种新的第三类靶向gp41的HIV融合抑制剂。提出了描述N36（Mut(e,g)）与预发夹中间体相互作用的定量模型。
The pre-hairpin intermediate of gp41 from the human immunodeficiency virus (HIV) is the target for two classes of fusion inhibitors that bind to the C-terminal region or the trimeric coiled-coil of N-terminal helices, thereby preventing formation of the fusogenic trimer of hairpins. Using rational design, two 36-residue peptides, N36(Mut(e,g)) and N36(Mut(a,d)), were derived from the parent N36 peptide comprising the N-terminal helix of the gp41 ectodomain (residues 546-581 of HIV-1 envelope), characterized by analytical ultracentrifugation and CD, and assessed for their ability to inhibit HIV fusion using a quantitative vaccinia virus-based fusion assay. N36(Mut(e,g)) contains nine amino acid substitutions designed to disrupt interactions with the C-terminal region of gp41 while preserving contacts governing the formation of the trimeric coiled-coil. N36(Mut(a,d)) contains nine substitutions designed to block formation of the trimeric coiled-coil but retains residues that interact with the C-terminal region of gp41. N36(Mut(a,d)) is monomeric, is largely random coil, does not interact with the C34 peptide derived from the C-terminal region of gp41 (residues 628-661), and does not inhibit fusion. The trimeric coiled-coil structure is therefore a prerequisite for interaction with the C-terminal region of gp41. N36(Mut(e,g)) forms a monodisperse, helical trimer in solution, does not interact with C34, and yet inhibits fusion about 50-fold more effectively than the parent N36 peptide (IC50 -similar to308 nM versus similar to16 muM). These results indicate that N36(Mut(e,g)) acts by disrupting the homotrimeric coiled-coil of N-terminal helices in the pre-hairpin intermediate to form heterotrimers. Thus N36(Mut(e,g)) represents a novel third class of gp41-targeted HIV fusion inhibitor. A quantitative model describing the interaction of N36(Mut(e,g)) with the pre-hairpin intermediate is presented.