Development of hybrid structural-MS to determine the 3D structures of protein complexes

开发混合结构 MS 以确定蛋白质复合物的 3D 结构

• 批准号: 1655397
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1655397
• 关键词: Development; hybrid; structural; MS; determine

Hydrogen deuterium exchange MS (HDX-MS) permits the real-time characterisation of protein dynamics and solvent accessibility on the peptide level. Recent advances in HDX-MS have turned this complex procedure into a robust, high-throughput and information rich tool. The aim of this studentship is to develop ways to extract information embedded in HDX patters for use with structural elucidation and dynamical characterisation of protein complexes. The student will develop ways to characterise native state dynamics from atomic structures that will then be used as a reference point to explain non-canonical protein motions involved in function and also to elucidate structural features such as binding interfaces. HDX-MS will also be used to interrogate the folding of protein complexes such as the integrity of binding interfaces and protein folds under mildly denaturing conditions that are typically used to generate sub-complexes. Where possible electron transfer dissociation will be used to access residue-specific information and to assess the utility of protection values in HDX-MS to map the free energy surfaces of proteins and their complexes.Labile protein protons exchange for bulk by rates determined by the chemical exchange factors and secondary structural effects. When the surrounding media is deuterium oxide these exchange events are coupled to a change in mass that can be tracked using high resolution mass spectrometry. HDXMS is important as is has the potential to allow the determination of the energy surface of a protein complex which is currently intractable by other methods. The student will be using HDXMS, high resolution MS, ion mobility mass spectrometry, native MS and potentially electron transfer dissociation. The project will have impact for Waters Corporation who design and Manufacture the instruments.

氢重离子交换质谱仪(HDX-MS)可以在多肽水平上实时表征蛋白质动力学和溶剂可及性。HDX-MS的最新进展已将这一复杂的过程转变为一种强大、高通量和信息丰富的工具。这个学生的目的是开发方法来提取嵌入在HDX模式中的信息，用于蛋白质复合体的结构阐明和动态表征。学生将开发方法来表征原子结构的本征状态动力学，然后将被用作参考点，以解释涉及功能的非正则蛋白质运动，并解释结构特征，如结合界面。HDX-MS还将用于询问蛋白质复合体的折叠，如结合界面和蛋白质折叠在温和变性条件下的完整性，这些条件通常用于生成子复合体。在可能的情况下，将使用电子转移解离来获取残基特定的信息，并评估HDX-MS中保护值的有效性，以绘制蛋白质及其络合物的自由能表面。不稳定的蛋白质质子交换体积的速率由化学交换因子和二级结构效应决定。当周围的介质是氧化氢时，这些交换事件与质量的变化有关，可以使用高分辨率质谱仪进行跟踪。HDXMS很重要，因为它有可能确定蛋白质复合体的能级表面，这是目前用其他方法难以解决的。学生将使用HDXMS、高分辨率MS、离子迁移质谱仪、自然MS和潜在的电子转移解离。该项目将对设计和制造这些仪器的Waters Corporation产生影响。