A screening assay for antiviral compounds targeted to the HIV-1 gp41 core structure using a conformation-specific monoclonal antibody
A screening assay for antiviral compounds targeted to the HIV-1 gp41 core structure using a conformation-specific monoclonal antibody
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DOI：
10.1016/s0166-0934(99)00041-5
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发表时间：
1999-06-01
影响因子：
3.1
通讯作者：
Debnath, AK
中科院分区：
文献类型：
--
作者：
Jiang, SB;Lin, K;Debnath, AK
人类免疫缺陷病毒1型（HIV-1）gp 41在病毒与靶细胞的膜融合中起重要作用。gp 41胞外域含有两个与N和C末端相邻的七肽重复区。来自这两个区域的肽，称为N肽和C肽，是HIV-1感染的有效抑制剂，并且可以彼此相互作用形成六链卷曲螺旋，代表gp 41的融合核心结构。产生单克隆抗体，命名为NC-1，其特异性结合由N和C-肽形成的复合物，但不结合单个肽。用NC-1建立了一种酶联免疫吸附试验（ELISA），用于检测N肽和C肽形成的复合物，并用于筛选可能干扰复合物形成和抑制HIV-1感染的抗病毒药物。C肽中的单点突变消除了复合物的形成，也消除了它们的抗HIV-1活性。发现一种苯偶氮萘磺酸衍生物，命名为ADS-J1，抑制NC-1可检测复合物的形成和HIV-1介导的膜融合，表明所述ELISA适用于快速筛选靶向HIV-1 gp 41核心结构的HIV-1抑制剂的有机化合物库。(C)1999 Elsevier Science B. V.保留所有权利。
The human immunodeficiency virus type 1 (HIV-1) gp41 plays an important role in membrane fusion between viruses and target cells. The gp41 ectodomain contains two heptad repeat regions adjacent to the N and C-termini. Peptides derived from these two regions, designated N and C-peptides, are potent inhibitors of HIV-1 infection and can interact with each other to form a six-stranded coiled-coil, representing the fusogenic core structure of gp41. A monoclonal antibody was generated, designated NC-1, which specifically binds to the complex formed by the N and C-peptides, but not to the individual peptides. An enzyme linked immunosorbent assay (ELISA) was developed using NC-1 for detecting complex formed by N and C-peptides and for screening of organic compounds for antiviral agents that may interfere with complex formation and inhibit HIV-1 infection. Single point mutations in the C-peptides abolish the complex formation also eliminate their anti-HIV-1 activity. A phenylazo-naphthalene sulfonic acid derivative, designated ADS-J1, was found to inhibit both formation of NC-1 detectable complex and HIV-1-mediated membrane fusion, suggesting that the described ELISA is applicable to rapid screening of libraries of organic compounds for HIV-1 inhibitors targeted to the HIV-1 gp41 core structure. (C) 1999 Elsevier Science B.V. All rights reserved.