Max-Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Protein Expression Laboratory, NIAMS, National Institutes of Health, Bethesda, MD, USA.
FEBS Lett. 2018 Mar;592(6):939-948. doi: 10.1002/1873-3468.13010. Epub 2018 Mar 5.
The HIV-1 envelope gp120/gp41 trimer mediates viral membrane fusion. After cluster of differentiation-4 recognition, gp120 detaches from the virus, exposing gp41 which triggers fusion. During the fusion process, gp41 may not remain trimeric, which could have functional importance. Here, we probe the reversible association of full length gp41 (minus the cytoplasmic domain) in detergent micelles (with probes attached to transmembrane domain) by fluorescence resonance energy transfer (FRET) with a μm dissociation constant. This is compared with other methods. A gp41-targeted fusion inhibitor must interfere with this transition, and monomeric, partially monomeric or trimeric states all present potential binding epitopes. The gp41 self-association is a valid drug target model and FRET, a potential high-throughput assay system, could be used to screen drug libraries.
HIV-1 包膜 gp120/gp41 三聚体介导病毒膜融合。在识别分化群-4 后，gp120 从病毒上脱离，暴露出 gp41，gp41 触发融合。在融合过程中，gp41 可能不再保持三聚体状态，这可能具有功能重要性。在这里，我们通过荧光共振能量转移（FRET）用探针（附着在跨膜域上）探测去污剂胶束中全长 gp41（减去细胞质域）的可逆缔合（μm 解离常数），并与其他方法进行比较。靶向 gp41 的融合抑制剂必须干扰这种转变，而单体、部分单体或三聚体状态都存在潜在的结合表位。gp41 自缔合是一个有效的药物靶标模型，FRET 是一种潜在的高通量检测系统，可用于筛选药物库。
