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

氢氘交换MS（HDX-MS）允许在肽水平上实时表征蛋白质动力学和溶剂可及性。HDX-MS的最新进展已经将这个复杂的过程变成了一个强大的，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，用于蛋白质复合物的结构阐明和动力学表征。学生将开发方法来从原子结构，然后将被用作一个参考点，以解释参与功能的非典型蛋白质运动，并阐明结构特征，如结合界面。HDX-MS还将用于询问蛋白质复合物的折叠，例如通常用于产生亚复合物的轻度变性条件下的结合界面和蛋白质折叠的完整性。在可能的情况下，电子转移解离将用于访问残基特异性信息，并评估HDX-MS中保护值的实用性，以绘制蛋白质及其复合物的自由能表面。当周围介质是氧化氘时，这些交换事件与质量变化相关联，可以使用高分辨率质谱法来跟踪质量变化。HDXMS是重要的，因为它有可能允许确定蛋白质复合物的能量表面，这是目前其他方法难以处理的。学生将使用HDXMS，高分辨率MS，离子迁移质谱，天然MS和潜在的电子转移解离。该项目将对设计和制造仪器的沃茨公司产生影响。

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.