Exploiting ion mobility and high performance mass spectrometry for in-depth characterization of neuropeptides in hypoxia and pH stress

在化学系化学测量与成像项目的支持下，威斯康星大学麦迪逊分校的李凌军教授正在开发一种新的基于质谱的技术来研究环境压力下神经肽的变化。李教授和她的团队专注于研究螃蟹在缺氧或酸性或碱性条件下（pH胁迫）的结构和神经肽浓度是如何变化的。由于神经肽是调节各种生理过程的最多样化和最基本的分子类别之一，因此需要新的方法和技术来更完整地描述这些信号分子并量化它们对环境应激反应的变化。李教授和她的团队开发了一套基于离子迁移率质谱的工具，以分离不同大小与电荷比的不同肽。对检测到的不同肽进行分类将有助于了解它们在大脑中的作用。这项资助的研究利用了分析化学工具的力量来推进神经科学研究。它为李教授小组的研究生和本科生提供了在化学和神经科学领域从事研究工作的良好培训机会。该项目旨在开发强大的离子迁移质谱（IM-MS）技术的新用途，以解决神经肽研究面临的几个剩余技术挑战。具体来说，李教授和她的团队获得资助，开发一种基于IM-MS耦合微分离，高性能MS和分子动力学模拟的新型多管齐下的方法，用于大规模发现，定量和神经肽的结构阐明。他们计划改进离子迁移辅助大规模肽组学分析和精确定量的方法，将其与各种微分离平台和质谱成像相结合，用于全面的神经肽分析。此外，他们将开发一种独特的基于IM-MS的策略来探测神经肽的新型翻译后修饰（PTMs），如D/ l肽外显子和o链糖基化。新的方法是提高对缺氧和pH胁迫的神经化学信号分析。这项提议所带来的技术进步将适用于包括人类在内的许多神经系统的生化和肽能信号的大规模分析。通过探索神经化学对应激反应的分子洞察力，可以更好地理解参与适应环境应激的神经内分泌调节机制，这一过程在所有生物体中都是高度保守的，以维持它们在面对外部和内部产生的“刺激”时的生存。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Lingjun Li at the University of Wisconsin, Madison, is developing new mass spectrometry-based technology to study neuropeptide changes involved in environmental stress. Professor Li and her team focus on the study of how the structures and the concentrations of neuropeptides in a crab change under the lack of oxygen (hypoxia) or under acidic or basic conditions (pH stress). As neuropeptides represent one of the most diverse and essential classes of molecules that regulate various physiological processes, new approaches and techniques are needed to enable a more complete description of these signaling molecules and quantify their changes in response to environmental stressors. Professor Li and her group develop a set of tools based on ion-mobility mass spectrometry to separate different peptides of different size-to-charge ratios. Cataloging different peptides detected will then help to understand their roles in the brain. The funded research takes advantage of the power of analytical chemistry tools to advance neuroscience research. It provides excellent training opportunities to graduate students and undergraduate students in Professor Li's group to pursue a research career at the interface of chemistry and neuroscience. This project seeks to exploit novel use of the powerful ion mobility mass spectrometry (IM-MS) technology to address several remaining technical challenges facing neuropeptide research. Specifically, Professor Li and her team are funded to develop a novel multi-pronged approach based on IM-MS coupling to microseparations, high-performance MS, and molecular dynamic simulations for large-scale discovery, quantitation and structural elucidation of neuropeptides. They plan to improve methodologies for ion mobility assisted large-scale peptidomic analysis and accurate quantitation, its coupling to various microseparation platforms and mass spectral imaging for comprehensive neuropeptide analysis. Furthermore, they will develop a unique IM-MS based strategies to probe novel posttranslational modifications (PTMs) of neuropeptides such as D/L-peptide epimers and O-linked glycosylation. The new methodology is to improve neurochemical signaling analysis in response to hypoxia and pH stress. The technological advancements resulting from this proposal will be applicable to large-scale analysis of biochemical and peptidergic signaling in many nervous systems, including humans. The molecular insights gained from probing neurochemistry in response to stress could lead to a better understanding of neuroendocrine regulatory mechanisms involved in adapting to environmental stress, a process that is highly conserved among all organisms to maintain their survival in the face of both externally and internally generated "stimuli".