Leveraging Ion Mobility and High Resolution Mass Spectrometry to Probe Neuropeptide Modifications in Environmental Stress

利用离子淌度和高分辨率质谱来探测环境应激中的神经肽修饰

• 批准号: 2108223
• 负责人: Lingjun Li
• 金额: $ 43.5万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2108223&HistoricalAwards=false
• 关键词: Leveraging; Ion; Mobility; Resolution; Mass

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Lingjun Li and her group at the University of Wisconsin-Madison are developing new analytical measurement tools to probe environmentally induced changes in neuropeptides. Specifically, the Li group seeks to exploit state-of-the-art analytical methods (mass spectrometry and ion mobility spectrometry) using chemical “tags” to enhance sensitivity and throughput during characterization and quantification of signaling peptides and their modifications in response to copper toxicity and hypoxia (oxygen deprivation) stress of cells. The technology being developed is expected to be transferable to the study of other biological systems. The molecular insights gained from the proposed studies have the potential to enhance understanding of normal and dysfunctional biology involving neuronal peptide signaling. This project provides excellent training opportunities for students who are interested in a career at the interface between chemical measurements and neuroscience. Dr. Li also works to bring awareness and participation in these research activities through partnership and outreach with high school internship programs that help to stimulate students’ interests in the STEM field at an early stage.The Li group seeks to develop an innovative analytical platform that integrates microseparations with ion mobility-mass spectrometry (IM-MS) and high-resolution MS to probe dynamic changes and structural effects of novel post-translational modifications of neuropeptides involved in copper toxicity, hypoxia, and hypercapnia. Specifically, Professor Li and her team are pursuing the following research goals: (1) to employ design-of-experiment algorithms and higher-plexing chemical tags to expand analytical throughput for neuropeptide quantitation; (2) to couple rapid capillary electrophoresis separation with IM-MS for comprehensive neuropeptide analysis in copper toxicity and hypoxia/hypercapnia; (3) to combine “collision-induced unfolding” IM-MS with surface plasmon resonance and molecular dynamics simulation for in-depth structural and conformational analysis of peptide epimers and their interaction with receptors; and (4) to construct a large-scale collision cross section (CCS) database for neuropeptide isoforms and glycosylated neuropeptides to facilitate functional discovery and evaluation of novel glycosylated neuropeptides. The research seeks to advance measurement science and accelerate its application to solve challenging biological questions regarding the functional roles of signaling peptides in the regulation and adaptation to heavy metal toxicity and hypoxia.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系化学测量和成像项目的支持下，威斯康星大学麦迪逊分校的Lingjun Li教授和她的团队正在开发新的分析测量工具，以探测环境引起的神经肽变化。 具体而言，Li小组寻求利用最先进的分析方法（质谱法和离子迁移谱法），使用化学“标签”来提高表征和定量信号肽及其对细胞铜毒性和缺氧（氧剥夺）应激的反应的修饰期间的灵敏度和通量。 正在开发的技术有望转移到其他生物系统的研究中。 从拟议的研究中获得的分子见解有可能增强对涉及神经元肽信号传导的正常和功能失调生物学的理解。 该项目为那些对化学测量和神经科学之间的界面感兴趣的学生提供了极好的培训机会。 李博士还致力于通过与高中实习计划的合作和推广，帮助激发学生在早期阶段对STEM领域的兴趣，从而提高人们对这些研究活动的认识和参与。李博士的团队旨在开发一个创新的分析平台，将微分离与离子迁移-质谱（IM-MS）和高分辨率MS相结合，以探索新的后处理过程的动态变化和结构效应。涉及铜毒性、缺氧和高碳酸血症的神经肽的翻译修饰。 具体而言，李教授及其团队正致力于以下研究目标：（1）采用实验设计算法和更复杂的化学标签来扩大神经肽定量的分析通量;（2）将快速毛细管电泳分离与IM-MS结合起来，用于铜中毒和缺氧/高碳酸血症的神经肽综合分析;（3）将快速毛细管电泳分离与IM-MS结合起来，用于铜中毒和缺氧/高碳酸血症的神经肽分析。（3）将联合收割机“碰撞诱导去折叠”IM-MS与表面等离子体共振和分子动力学模拟相结合，对肽差向异构体及其与受体的相互作用进行深入的结构和构象分析;（4）构建神经肽异构体和糖基化神经肽的大规模碰撞截面（CCS）数据库，以促进新型糖基化神经肽的功能发现和评价。 该研究旨在推进测量科学，并加速其应用，以解决具有挑战性的生物学问题，即信号肽在调节和适应重金属毒性和缺氧中的功能作用。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

HyPep: An Open-Source Software for Identification and Discovery of Neuropeptides Using Sequence Homology Search.

• DOI: 10.1021/acs.jproteome.2c00597
• 发表时间: 2023-02-03
• 期刊: JOURNAL OF PROTEOME RESEARCH
• 影响因子: 4.4
• 作者: Vu, Nhu Q.;Yen, Hsu-Ching;Fields, Lauren;Cao, Weifeng;Li, Lingjun
• 通讯作者: Li, Lingjun

Enrichment and fragmentation approaches for enhanced detection and characterization of endogenous glycosylated neuropeptides.

• DOI: 10.1002/pmic.202100375
• 发表时间: 2023-02
• 期刊: Proteomics
• 影响因子: 3.4

Definitive Screening Designs to Optimize Library-Free DIA-MS Identification and Quantification of Neuropeptides.

确定的筛选设计，以优化无文库的DIA-MS识别和神经肽的定量。

• DOI: 10.1021/acs.jproteome.3c00088
• 发表时间: 2023-05-05
• 期刊: JOURNAL OF PROTEOME RESEARCH
• 影响因子: 4.4
• 作者: Phetsanthad, Ashley;Carr, Austin V.;Fields, Lauren;Li, Lingjun
• 通讯作者: Li, Lingjun

Multi-Faceted Mass Spectrometric Investigation of Neuropeptides in Callinectes sapidus.

• DOI: 10.3791/63322
• 发表时间: 2022-05-31
• 期刊: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
• 影响因子: 1.2
• 作者: Phetsanthad, Ashley;Vu, Nhu Q.;Li, Lingjun
• 通讯作者: Li, Lingjun

Bicarbonate buffers can promote crosslinking and alternative gas-phase dissociation pathways for multiprotein complexes

• DOI: 10.1016/j.ijms.2021.116687
• 发表时间: 2021-09-01
• 期刊: INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
• 影响因子: 1.8
• 作者: Li，Gongyu;Huang，Junfeng;Ruotolo，Brandon T.
• 通讯作者: Ruotolo，Brandon T.