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。
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.
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.
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.
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.
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.
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.
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.
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.