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

氢氘交换质谱（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的最新进展已将这一复杂的程序转变为强大、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模式的信息，用于蛋白质复合物的结构阐明和动态特性。学生将开发从原子结构中表征自然状态动力学的方法，然后将其用作解释涉及功能的非规范蛋白质运动的参考点，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，例如结合界面的完整性和通常用于生成亚复合物的轻度变性条件下的蛋白质折叠。在可能的情况下，电子转移解离将用于获取残基特异性信息，并评估HDX-MS中保护值的效用，以绘制蛋白质及其复合物的自由能表面。不稳定蛋白质子的交换速率由化学交换因子和次级结构效应决定。当周围介质是氧化氘时，这些交换事件与质量变化相耦合，可以使用高分辨率质谱法进行跟踪。HDXMS很重要，因为它有潜力确定蛋白质复合物的能量表面，这是目前其他方法难以解决的问题。学生将使用HDXMS，高分辨率MS，离子迁移质谱，原生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特斯公司产生影响。

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.