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NMR assignments of vaccinia virus protein A28: an entry-fusion complex component
Biomolecular NMR Assignments ( IF 0.9 ) Pub Date : 2021-01-04 , DOI: 10.1007/s12104-020-09993-0
Danni Wu, Yuan-Chao Lou, Wen Chang, Der-Lii M. Tzou

Vaccinia virus (VACV) belonging to the poxvirus family enters the host cell via two different entry pathways; either endocytosis or virus/host cell membrane fusion. With respect to the virus/host cell membrane fusion, there are eleven viral membrane proteins forming a complicated entry-fusion complex (EFC), including A28, A21, A16, F9, G9, G3, H2, J5, L5, L1 and O3, to conduct the fusion function. These EFC components are highly conserved in all poxviruses and each of them is essential and necessary for the fusion activity. So far, with the exceptions of L1 and F9 whose crystal structures were reported, the structural information about other EFC components remains largely unclear. We aim to conduct a structural and functional investigation of VACV virus-entry membrane protein A28. In this work, we expressed and purified a truncated form of A28 (14 kDa; residues 38–146, abbreviated as tA28 hereinafter), with deletion of its transmembrane domain (residues 1–22) and a hydrophobic segment (residues 23–37). And the assignments of its backbone and side chain 1H, 13C and 15N chemical shifts of tA28 are reported. The secondary structure propensity from TALOS+ indicates that tA28 does contain three α-helices, six β-strands and connecting loops. Aside from this, we demonstrated that tA28 does interact with fusion suppressor viral protein A26 (residues 351–500) by the 1H–15N HSQC spectrum. We interpret that A28 binding to A26 deactivates EFC fusion activity. The current study provides a valuable framework towards further structural analyses of this protein and for better understanding virus/host cell membrane fusion mechanism in association with virus entry.



中文翻译:

痘苗病毒蛋白 A28 的 NMR 分配:一种进入融合复合物组分

属于痘病毒家族的痘苗病毒 (VACV) 通过两种不同的进入途径进入宿主细胞;内吞作用或病毒/宿主细胞膜融合。在病毒/宿主细胞膜融合方面,有11种病毒膜蛋白形成复杂的进入融合复合物(EFC),包括A28、A21、A16、F9、G9、G3、H2、J5、L5、L1和O3 ,进行融合功能。这些 EFC 成分在所有痘病毒中都是高度保守的,它们中的每一个对于融合活动都是必不可少的。到目前为止,除了 L1 和 F9 的晶体结构已被报道,其他 EFC 组分的结构信息仍不清楚。我们的目标是对 VACV 病毒进入膜蛋白 A28 进行结构和功能研究。在这项工作中,我们表达并纯化了截短形式的 A28(14 kDa;残基 38-146,以下缩写为 tA28),删除其跨膜结构域(残基 1-22)和疏水片段(残基 23-37)。以及其主链和侧链的分配报道了tA28的1 H、13 C和15 N化学位移。来自 TALOS+ 的二级结构倾向表明 tA28 确实包含三个 α-螺旋、六个 β-链和连接环。除此之外,我们通过1 H– 15 N HSQC 谱证明了 tA28 确实与融合抑制病毒蛋白 A26(残基 351–500)相互作用。我们认为 A28 与 A26 的结合使 EFC 融合活性失活。目前的研究为进一步对该蛋白的结构分析和更好地理解与病毒进入相关的病毒/宿主细胞膜融合机制提供了一个有价值的框架。

更新日期:2021-01-05
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