开发病毒疫苗和抗病毒药物极具挑战性。 病毒非常复杂,以至于研究人员首先需要解析病毒结构并了解其生命周期。这些知识可以成为有效的开发疫苗的基础和寻找可以抑制或阻断病毒生命周期中关键生命进程的药物提供帮助。 您是否正在被病毒研究的复杂性所困扰?欢迎咨询了解 NanoTemper 技术如何为您的科研助力。

病毒结构和功能

了解病毒的构成——核酸,蛋白质衣壳,还有些含有脂质包膜,但只知道这些是远远不够的。研究人员仍然需要弄清楚这些成分是如何有机组合在一起发挥功能的,比如利用电子显微镜观察晶体结构,以及通过研究与宿主细胞蛋白质和核酸的相互作用来了解上述成分在病毒生命周期中发挥什么作用。

 


 

从MERS-CoV Nsp15的结构和功能研究中获取药物开发线索

Nsp15 plays an essential role in the life cycle of coronavirus (CoV). But the structural information of this protein from MERS-CoV is missing. To shed light on its structure and functionality, this study looked at the complex formation between Nsp15 and other non-structural viral proteins. MST was used to confirm that Nsp15 associates to Nsp8 and Nsp8/Nsp7 with low micromolar affinities, and looked at how this association might affect catalytic activity.

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探索 CoV Nsp9 二聚体的形成及其对核酸结合亲和力的影响

Nsp9 is an important RNA binding subunit in the RNA-synthesizing machinery of all CoV. Understanding the mechanism of nsp9 dimerization and nucleic acid binding provides new insight for antiviral drug development. The authors used MST to get the binding affinity (Kd) of nsp9 with various mutations and their effects in dimerization and binding to ssDNA – while EMSA could only confirm binding but not measure the affinity.

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帮助解析复杂的刺突糖蛋白的冷冻电镜结构

The entry of CoV into cells is mediated by the transmembrane spike glycoprotein S, which forms a trimer and has receptor-binding and membrane fusion functions. Understanding the structure of this glycoprotein pre- and post-fusion can help in the design of vaccines. MST revealed that the binding affinity between glycoprotein S from mouse hepatitis virus and the soluble mouse receptor was in the nanomolar range.

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Monitor RNA release from viral capsids

RNA release is a critical step during the viral infection process. This group used nanoDSF to examine subtle differences in full vs. empty capsids to characterize the uncoating of a picornavirus. They could also use this information to determine how the stabilization of a viral capsid with a small molecule helps prevent uncoating, and thus infection.

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See how viral uncoating relates to infectivity

The mechanism of uncoating is important for viral infectivity, but many aspects of the process remain unclear. When HIV-1 undergoes the uncoating process, it sheds capsid proteins from its core. See how one group used Tycho to measure how mutations to the capsid proteins affected the stability of the shedded proteins, and therefore impacted infectivity.

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Virology studies can be used to expand the field of gene therapy. See how AAVs and other vectors are being used to target difficult-to-treat diseases.

 

See gene therapy spotlight

病毒生命周期

 

为了自我复制,病毒必须严格遵循一个多步骤的生物学过程。它始于对宿主细胞膜上受体的附着,并随着病毒后代的释放而结束。其中一种方法科研人员可以用来持续研究病毒复制过程中的每一个重要的生物学步骤,就是通过观察和检测参与此过程的蛋白质之间是如何相互作用的。

 


 

用多价抑制剂阻断甲型流感病毒的进入

Influenza A virus spike protein hemagglutinin binds to sialic acid on the cell membrane in a multivalent way. Designing multivalent binders is a promising approach to prevent infection. This study presents a multivalent binder that is shown to inhibit virus infection in vitro, ex vivo, and in vivo. MST is used to validate the optimal construction of the inhibitor measuring its interactions with Influenza A virus.

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揭示阻止 HIV-1 物种特异性感染的因素

Cellular protein TRIM5α gives resistance to HIV-1 in rhesus monkeys, but not in humans. It binds to the virus protein shell despite the high mutation rate seen in many retroviruses. This study sought to find out what virus capsid arrangements were responsible for the species-specific resistance. MST measurements revealed that the HIV-1 capsid surface is critical for the binding of TRIM5α and its species-specific protection against infection in rhesus monkeys.

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Learn what binding affinity studies can tell you about viruses and how you can defeat them

 

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In the past decade, my research has focused on characterizing multivalent binders against the spike proteins of the influenza A virus. MST made it possible to determine binding constants using whole virus particles, which revealed important insights on multivalent binders interacting with the native virus surface.

我强烈推荐这项优秀的技术用来做病毒结合方面的研究。

柏林自由大学
丹尼尔·劳斯特博士

 

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抗病毒药

抗病毒药物用于治疗病毒感染,并被开发为可以阻断病毒生命周期的特定步骤。 这些药物通过与特定分子结合而发挥作用,因此它们可以干扰病毒与其受体,复制或病毒装配的附着。

 


 

在 5% DMSO 条件下筛选衣壳装配抑制剂

Feline immunodeficiency virus (FIV) is a retrovirus that causes AIDS in cats. There is a lack of antiretroviral drugs for FIV infections mostly because of the little structural information available for FIV proteins. This study screened 400 compounds, looking for inhibitors of the virus capsid assembly in vitro. With MST the authors could screen in the presence of 5% DMSO, resolving the solubility problems they encountered for some compounds. Another advantage was that MST used only nanomolar amounts of protein.

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让研发始终领先于丙型肝炎病毒基因的遗传变异

The high genetic variability of hepatitis C virus (HCV) and rapid development of drug-resistant strains are driving the search for new direct-acting antiviral agents. In this publication, the authors focused on agents that target the HCV protein NS5A. Their goal was to determine how they interacted and how they affected its function. With MST they revealed that two clinical relevant inhibitors bound tightly to NS5A and inhibited binding to RNA. They also showed that these inhibitors do not affect NS5A dimerization.

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