IDBR: Type A. High-Performance Surface-induced Dissociation MS/MS

俄亥俄州州立大学获得了一个奖项，以开发和仪器，可以通过改进质谱仪中复合物的解离来表征蛋白质复合物。生命系统中的蛋白质可以发挥结构作用，例如皮肤和头发中的蛋白质;它们也可以发挥积极作用，例如帮助消化的酶，或携带氧气和二氧化碳通过身体的血红蛋白。细胞过程如代谢、细胞信号传导、基因表达、运输、细胞周期调节和亚细胞结构的形成取决于蛋白质复合物/组装体的形成和动态相互作用。这些蛋白质复合物也可以与其他生物分子（DNA/RNA）结合，以特定的方式相互作用。由于蛋白质结构与功能密切相关，因此需要有效的工具来表征蛋白质-蛋白质和蛋白质-配体复合物。拟议的研究将导致表面诱导解离装置集成到多种类型的质谱仪中，这些质谱仪将提供有关生物学中重要的蛋白质复合物结构的信息。 该项目的发现将在大学课程和国际，国家，区域和地方的质谱和生物分子结构研讨会/会议上以及同行评审文献中展示。 本科生和研究生以及博士后助理将接受仪器开发方面的培训，并将通过与私营部门合作伙伴和当地电子和机械商店的互动获得协作技能。 早期的SID采用者将作为修改成功的直接共鸣板。该项目是供应商和平台中立的，直接包括多个私营部门合作伙伴，阿尔达拉技术，沃茨（Q-IM-TOF，四极离子迁移率飞行时间），Thermo（Orbitrap）和Bruker（ICR，离子回旋共振），为项目团队提供了在公司用户会议上发言的机会，并为这些合作伙伴提供了评估仪器修改的商业价值的机会。将在高/超高分辨率质谱仪中开发表面诱导解离（SID），并改进其在离子迁移率（IM）质谱仪中的应用，其目的是表征蛋白质复合物。这项研究将使许多不同领域的研究人员对蛋白质复合物的拓扑结构表征感兴趣。对Thermo和Bruker的计划性合作访问是研究的必要组成部分，以熟悉所需的仪器平台，计划修改，并允许供应商评估进展。 最终的仪器配置将结合联合收割机IM或高/低分辨能力与有效的SID碎裂和增强的产物离子收集。 所提出的组合目前在任何商业仪器上都是不可用的，预计将提高分析质量，并扩大可通过质谱法（MS）有效表征的生物样品的范围。与过去使用的仅DC聚焦相比，SID的计划改进利用了碰撞后离开表面的离子的RF聚焦。将SID纳入目前的商业超高分辨率平台尚未完成，预计将大大提高生物系统分析的范围和深度。 在布鲁克仪器中，这种改进将使SID不仅作为一种可能的额外活化方法，而且作为碰撞诱导解离（CID）的替代品。该项目的整体智力价值是高性能质谱仪的适应，包括一个更高性能的活化方法，用于难降解分析物的碎片，现有的方法无法提供直接的子结构信息。

An award is made to Ohio State University to develop and instrument that can characterize protein complexes through improved dissociation of the complexes in mass spectrometers. Proteins in living systems can play structural roles, such as those in skin and hair; they can also play active roles such as enzymes that help with digestion, or hemoglobin that carries oxygen and carbon dioxide through the body. Cellular processes such as metabolism, cell signaling, gene expression, trafficking, cell cycle regulation, and the formation of subcellular structures are contingent upon the formation and dynamic interactions of complexes/assemblies of proteins. These protein complexes may also be bound to other biomolecules (DNA/RNA), interacting in specific ways. Because protein structure is tied to function, effective tools are needed to characterize protein-protein and protein-ligand complexes. The proposed research will result in integration of surface-induced dissociation devices into multiple types of mass spectrometers that will provide information on the structures of protein complexes important in biology. Discoveries from the project will be presented in university courses and at international, national, regional, and local workshops/ conferences on mass spectrometry and biomolecule structure and in the peer-reviewed literature. The undergraduate and graduate students and the postdoctoral associate will be trained in instrument development and will also gain collaboration skills through interactions with the private sector partners and local electronics and machine shops. Early SID adopters will serve as a direct sounding board for the success of modifications. The project is vendor- and platform-neutral, with direct inclusion of multiple private sector partners, Ardara Technologies, Waters (Q-IM-TOF, quadrupole ion mobility time of flight), Thermo (Orbitrap), and Bruker (ICR, ion cyclotron resonance), providing the project team an opportunity to present at company user meetings and an opportunity for those partners to assess the commercial value of the instrument modification.This research will develop surface-induced dissociation (SID) in high/ultra-high resolution mass spectrometers and refine its use in an ion mobility (IM) mass spectrometer whose purpose is to characterize protein complexes. The proposed research will benefit researchers from many different fields interested in the characterization of the topology of protein complexes. Planned, collaborative visits to Thermo and Bruker are a necessary part of the research, to gain familiarity with needed instrument platforms, to plan modifications, and to allow the vendors to assess progress. The final instrument configurations will combine either IM or high/ultrahigh resolving power with efficient SID fragmentation and enhanced collection of product ions. The proposed combinations are currently unavailable on any commercial instrument and are expected to enhance the quality of analysis and expand the range of biological samples that can be effectively characterized by mass spectrometry (MS). Planned improvements to SID utilize RF focusing of the ions departing the surface post collision, in contrast with the DC-only focusing that has been used in the past. Inclusion of SID in current commercial ultra-high resolution platforms has not yet been accomplished and is expected to dramatically enhance both the scope and depth of analysis on biological systems. In the Bruker instrument, the modification will allow SID to be explored, not only as a possible additional activation method, but as a replacement for collision-induced dissociation (CID). The overall intellectual merit of the project is adaptation of high performance mass spectrometers to include a higher performance activation method for fragmentation of refractory analytes, for which existing methods fail to provide direct substructure information.