Biophysical studies of HIV-1 glycoprotein-41 interactions with peptides and small molecules - Effect of lipids and detergents.
Biophysical studies of HIV-1 glycoprotein-41 interactions with peptides and small molecules - Effect of lipids and detergents.
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DOI：
10.1016/j.bbagen.2020.129724
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发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Gochin M
中科院分区：
文献类型：
--
作者：
Zhou G;Chu S;Kohli A;Szoka FC;Gochin M
HIV-1糖蛋白-41胞外域的疏水口袋（HP）由介导6 HB形成的蛋白-蛋白界面内的两条N-七肽重复三聚体链定义。它是病毒融合抑制剂的潜在靶点，但其疏水性和与膜的原位接近性妨碍了对抑制剂相互作用的快速分析。我们评估了19 F核磁共振和荧光检测肽和小分子与HP结合的灵敏度，并探索了非变性去污剂或磷脂作为共溶剂的影响以及gp 41周围膜环境的潜在模拟物。芳香族氟的化学位移被发现是敏感的氢键网络发生的变化时，抑制剂从溶剂过渡到HP或到有序的洗涤剂胶束。自发荧光化合物的荧光强度和发射最大值响应于当地环境的变化。Gp 41-配体结合发生在所有条件下，但在洗涤剂的存在下减少。核磁共振和荧光研究表明，十二烷基磷酸胆碱（DPC）是一个贫穷的替代膜在这个系统中，而脂质体可以模拟膜环境。我们的研究结果表明，HP的高效小分子结合剂的开发可能会受到HP结合和双层膜结合之间的竞争的阻碍。
The hydrophobic pocket (HP) of HIV-1 glycoprotein-41 ectodomain is defined by two chains of the N-heptad repeat trimer, within the protein-protein interface that mediates 6HB formation. It is a potential target for inhibitors of viral fusion, but its hydrophobic nature and proximity to membrane in situ has precluded ready analysis of inhibitor interactions. We evaluated the sensitivity of 19F NMR and fluorescence for detecting peptide and small molecule binding to the HP and explored the effect of non-denaturing detergent or phospholipid as cosolvents and potential mimics of the membrane environment surrounding gp41. Chemical shifts of aromatic fluorines were found to be sensitive to changes in the hydrogen bonding network that occurred when inhibitors transitioned from solvent into the HP or into ordered detergent micelles. Fluorescence intensities and emission maxima of autofluorescent compounds responded to changes in the local environment. Gp41 - ligand binding occurred under all conditions, but was diminished in the presence of detergents. NMR and fluorescence studies revealed that dodecylphosphocholine (DPC) was a poor substitute for membrane in this system, while liposomes could mimic the membrane surroundings. Our findings suggest that development of high potency small molecule binders to the HP may be frustrated by competition between binding to the HP and binding to the bilayer membrane.
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影响因子：
3.4
作者：
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通讯作者：
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影响因子：
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作者：
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DOI：
10.1073/pnas.95.26.15613
发表时间：
1998-12-22
影响因子：
11.1
作者：
Chan, DC;Chutkowski, CT;Kim, PS
通讯作者：
Kim, PS