Oligomerization analysis as a tool to elucidate the mechanism of EBV latent membrane protein 1 inhibition by pentamidine
Oligomerization analysis as a tool to elucidate the mechanism of EBV latent membrane protein 1 inhibition by pentamidine
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DOI：
10.1016/j.bbamem.2020.183380
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发表时间：
2020-10-01
影响因子：
3.4
通讯作者：
Mineev, Konstantin S.
中科院分区：
文献类型：
--
作者：
Kot, Erik F.;Wang, Yibo;Mineev, Konstantin S.
潜伏膜蛋白1(LMP1)是爱泼斯坦-巴尔病毒(EBV)的基因产物，EBV是一种广泛传播的病毒，存在于全球90-95%的人口中。EBV可导致多种恶性肿瘤，LMP1被证明在其中起关键作用。LMP1仅以低聚形式存在，其第五跨膜结构域(TMD-5)对寡聚反应至关重要，D150被认为是LMP1激活的关键残基。在这里，我们提出了一种基于核磁共振的方法来处理具有缓慢构象交换的复杂齐聚平衡。用这种方法研究了DPC胶束中的TMD-5。我们证明了D150的pKa等于7.4。未带电形式的TMD-5结合成二聚体和三聚体，D150的去质子化诱导蛋白质的高阶齐聚并显著增强其三聚化。五烷胺主要与带电的TMD-5相互作用，破坏齐聚物，稳定单体和三聚体。通过计算机模拟，我们研究了TMD-5/五烷胺相互作用的结构基础。我们的数据表明，在自然条件下，D150可能在全长LMP1中充电。
Latent membrane protein 1 (LMP1) is a gene product of the Epstein-Barr virus (EBV), a widely spread virus present in 90-95% of the world's population. EBV can lead to several malignancies, in which LMP1 was shown to play a key role. LMP1 is active only in the oligomeric form and its fifth transmembrane domain (TMD-5) is critical for the oligomerization, with D150 identified as a key residue for LMP1 activation. Here we propose an NMR-based approach to treat the complex oligomerization equilibria with slow conformational exchange. Using this method we investigate the TMD-5 in DPC micelles. We show that the pKa of D150 equals 7.4. Uncharged form of TMD-5 associates into dimers and trimers, deprotonation of D150 induces the high-order oligomerization of the protein and enhances dramatically its trimerization. Pentamidine interacts mainly with the charged TMD-5, destroying the oligomers and stabilizing the monomer and trimer. Using computer simulations we investigate the structural basis of TMD-5/pentamidine interaction. Our data suggest that D150 is likely charged in the full-length LMP1 under native conditions.