SOLUTION SCATTERING STUDY ON PRO-FUSION INTERMEDIATE OF INFLUENZA HEMAGGLUTININ

流感血凝素促融合中间体的溶液散射研究

• 批准号: 7954929
• 负责人: HIROTSUGU TSURUTA
• 金额: $ 0.87万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954929
• 关键词: Biophysics; Cell-Matrix Junction; Computer Retrieval of Information on Scientific Projects Database; Data Quality; Endocytosis; Environment; Evolution; Funding; Future; Glycoproteins; Grant; Hemagglutinin; Influenza Hemagglutinin; Institution; Membrane Fusion; Molecular Conformation; Monitor; Nature; Preparation; Protocols documentation; Research; Research Personnel; Resources; Sampling; Solutions; Source; Time; United States National Institutes of Health; Variant; Virus; X-Ray Crystallography; improved; research study; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We study the pH dependent conformational switching of bromelain-released influenza hemagglutinin by small angle solution x-ray scattering. The trimeric hemagglutinin (HA) glycoprotein is responsible for both host cell attachment and membrane fusion in influenze virus. The acidic environment in endocytosis is known to trigger a large scale conformational switching. X-ray crystallography has given the three dimensional structures of the initial and the post fusion states. There is little structural information available so far for activated intermediate states due to their transient nature central to their host attachment and fusion functions. We trigger the conformational switching by lowering solution pH and monitor the evolution of HA conformation via solution x-ray scattering as a function of time over several minutes. We obtained encouraging results at BioCAT fairly similar to what we had obtained at SSRL in summer 2007. The improved data quality was evident at BioCAT. We observed x-ray scattering curves which has slightly different features at intermediary scattering angles than had been observed in 2007, indicating the slight variation in sample preparation. We are evaluating sample preparation protocol and planning on conducting additional experiments in near future.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们通过小角度溶液 X 射线散射研究了菠萝蛋白酶释放的流感血凝素的 pH 依赖性构象转换。三聚体血凝素 (HA) 糖蛋白负责流感病毒中的宿主细胞附着和膜融合。众所周知，内吞作用中的酸性环境会引发大规模的构象转换。 X射线晶体学给出了初始和融合后状态的三维结构。由于其瞬态性质对于宿主附着和融合功能至关重要，因此迄今为止对于激活的中间态几乎没有可用的结构信息。 我们通过降低溶液 pH 值来触发构象转换，并通过溶液 X 射线散射监测 HA 构象随时间变化的演变（几分钟内）。 我们在 BioCAT 上获得的令人鼓舞的结果与 2007 年夏季在 SSRL 上获得的结果非常相似。BioCAT 上数据质量的提高是显而易见的。我们观察到的 X 射线散射曲线在中间散射角处的特征与 2007 年观察到的略有不同，表明样品制备中存在轻微变化。我们正在评估样品制备方案并计划在不久的将来进行更多实验。