Departamento de Bioquímica e Química Orgânica, Instituto de Química, UNESP -Univ Estadual Paulista, 14800-900, Araraquara, SP, Brazil.
Instituto de Física de São Carlos, Universidade de São Paulo, USP, 13560-970, São Carlos, São Paulo, Brazil.
Virology. 2021 May;557:62-69. doi: 10.1016/j.virol.2021.02.012. Epub 2021 Feb 26.
Dengue virus infection depends on its fusion with the host membrane, where the binding occurs through interaction between proteins on the virus cell surface and specific viral receptors on target membranes. This process is mediated by the fusion peptide located between residues 98 and 112 (DRGWGNGCGLFGKGG) that forms a loop in domain II of dengue E glycoprotein. In this study, we evaluated the role of fusion peptide surrounding regions (88-97 and 113-123) of the Dengue 2 subtype on its interaction with the membrane and fusion activity. These sequences are important to stabilize the fusion peptide loop and increase fusion activity. Three peptides, besides the fusion peptide, were synthesized by SPPS using the Fmoc chemical approach. The first contains the fusion peptide and the C-terminal region of the loop (sequence 98-123); another contains the N-terminal region (88-112) and the larger peptide contains both regions (88-123). The peptides were able to interact with a model membrane. Differences in morphology of the monolayer promoted by the peptides were assessed by Brewster Angle Microscopy (BAM). Our data indicated that the C-terminal region of fusion peptide loop is more efficient in promoting fusion and interacting with the membrane than the N-terminal sequence, which is responsible for the electrostatic initial interaction. We propose a 2-step mechanism for the interaction of the dengue virus fusion peptide with the host membrane, where the N-terminal sequence docks electrostatically on the headgroups and then the C-terminal interacts via hydrophobic forces in the acyl chains.
登革热病毒感染依赖于其与宿主膜的融合,其中结合通过病毒细胞表面上的蛋白质与靶膜上的特定病毒受体之间的相互作用发生。该过程由位于残基 98 和 112 之间的融合肽(DRGWGNGCGLFGKGG)介导,该融合肽在登革热 E 糖蛋白的结构域 II 中形成环。在这项研究中,我们评估了登革热 2 型亚型融合肽周围区域(88-97 和 113-123)对其与膜相互作用和融合活性的作用。这些序列对于稳定融合肽环并提高融合活性很重要。除了融合肽之外,还使用 Fmoc 化学方法通过 SPPS 合成了三个肽。第一个肽包含融合肽和环的 C 末端区域(序列 98-123);另一个包含 N 末端区域(88-112),而较大的肽包含两个区域(88-123)。这些肽能够与模型膜相互作用。通过布鲁斯特角显微镜(BAM)评估肽促进的单层形态的差异。我们的数据表明,融合肽环的 C 末端区域比负责静电初始相互作用的 N 末端序列更有效地促进融合和与膜相互作用。我们提出了登革热病毒融合肽与宿主膜相互作用的两步机制,其中 N 末端序列静电结合在头基上,然后 C 末端通过酰基链中的疏水力相互作用。