Assessing the Biomolecular Structures that Result from Electrospray Ionization

评估电喷雾电离产生的生物分子结构

• 批准号: 10712440
• 负责人: Elyssia S Gallagher
• 金额: $ 33.64万
• 依托单位: BAYLOR UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712440
• 关键词: Address; Affect; Binding; Biological; Biological Process; Carbohydrates; Cells; Cellular biology; Charge; Complex; Development; Disease; Disease Progression; Electrospray Ionization; Exhibits; Future; Gases; Glycoconjugates; Goals; Infection; Ions; Isomerism; Ligand Binding; Mass Spectrum Analysis; Metals; Methodology; Methods; Modeling; Molecular; Phase; Polymers; Polysaccharides; Process; Property; Proteins; Research; Research Personnel; Signal Transduction; Sodium; Spectrometry, Mass, Electrospray Ionization; Structure; Techniques; Validation; Work; adduct; analytical method; deprotonation; host-microbe interactions; improved; insight; ion mobility; ionization; ionization technique; molecular dynamics; molecular recognition; novel; pathogen; protein complex; protein structure; protonation; tandem mass spectrometry; tool

Project Summary / Abstract Glycans, heterogeneous polymers of carbohydrates, interact with proteins to initiate a multitude of biologi- cal processes, including molecular recognition, cellular signaling, and host-microbe interactions. Mass spec- trometry (MS) methods have become powerful tools for characterizing the structures of glycans and their inter- actions with proteins. Electrospray ionization (ESI) is a common ionization method for transferring these bio- molecules from solution to the gas-phase for MS analysis. However, both glycans and proteins exhibit struc- tural changes during ESI. Thus, there is a critical need to understand how biomolecular structures are modified during and after the ESI process to determine (1) how researchers can deduce solvated structures from these techniques and (2) how analytical methods can be improved. To address this challenge, the Gallagher lab uses a combination of molecular dynamics (MD) simulations and MS methods to develop a fundamental, molecular perspective of ESI. The Gallagher lab performs MD simulations of ESI to observe carbohydrate ionization using metal ions as charge carriers. However, to more accurately model protein ionization, the Gallagher lab is developing methods to simulate protonation during ESI. These methods will be further developed to examine analyte ionization by deprotonation in negative-ion mode, and then applied to achieve a molecular perspective on the ionization of glycans and proteins. In paral- lel to the MD simulations, the Gallagher lab will perform MS analysis of metal-adducted glycans. Glycans readily ionize by coordinating to metal ions, with different metal-adducts enabling isomeric differentiation in both tandem MS and ion mobility-MS. However, glycans are often analyzed as sodium adducts because so- dium is a ubiquitous contaminant. The Gallagher lab is performing systematic studies examining the relation- ship between metal-ion properties and glycan characterization by MS. Finally, the Gallagher lab is applying native MS to characterize protein complexes. In native MS, noncovalent interactions are maintained in the gas phase; yet past work has suggested that proteins analyzed as positive versus negative ions have differ- ences in gas-phase stability. The Gallagher lab will examine how charging in ESI to form either positive or neg- ative ions is related to gas-phase protein structure, stability, and ligand-binding interactions. The overarching goal of the Gallagher lab is to develop and apply novel methodologies to characterize gly- cans, proteins, and their binding interactions. The research described in this proposal is significant because it will provide a fundamental perspective on ESI-MS and the molecular insights from this research will enable the rational development of future ESI-MS methods for characterizing these molecules. Ultimately, this work will facilitate the analysis of glycans, glycoconjugates, and protein complexes in cell biology and disease states, enabling these methods to be applied to address important biological hypotheses.

项目摘要 /摘要 碳水化合物的异质聚合物聚糖与蛋白质相互作用，以引发多种生物学 CAL过程，包括分子识别，细胞信号传导和宿主 - 微叶相互作用。质量规格 - TROTERITRY（MS）方法已成为表征聚糖及其相互作用的结构的强大工具 蛋白质的作用。电喷雾电离（ESI）是转移这些生物的常见电离方法 从溶液到气相的分子进行MS分析。然而，聚糖和蛋白质都表现出结构 ESI期间的刺激变化。因此，迫切需要了解生物分子结构如何修饰 在ESI过程中和之后，确定（1）研究人员如何从这些过程中推断出溶剂化的结构 技术和（2）如何改善分析方法。 为了应对这一挑战，Gallagher实验室结合了分子动力学（MD）模拟 和MS方法来开发ESI的基本，分子观点。 Gallagher实验室执行MD 使用金属离子作为电荷载体观察碳水化合物电离的模拟。但是，更多 精确模拟蛋白质电离，Gallagher Lab正在开发模拟质子化的方法 Esi。这些方法将进一步开发以检查通过负离子中去质子化的分析物电离 模式，然后应用以实现有关聚糖和蛋白质电离的分子观点。在 LEL到MD模拟，Gallagher Lab将对金属添加的聚糖进行MS分析。聚糖 通过与金属离子进行协调，很容易与不同的金属添加物进行电离 串联MS和离子迁移率MS。但是，通常将聚糖分析为钠加合物，因为 dium是一种普遍存在的污染物。 Gallagher实验室正在进行系统研究，以检查关系 - 在金属离子特性和MS的聚糖表征之间发货。最后，加拉格尔实验室正在申请 天然MS以表征蛋白质复合物。在天然MS中，在 气相；然而，过去的工作表明，分析为阳性与负离子的蛋白质有不同 在气相稳定性方面进行加密。 Gallagher实验室将检查ESI中的充电方式如何形成正面或负面 ATIVE离子与气相蛋白结构，稳定性和配体结合相互作用有关。 Gallagher实验室的总体目标是开发和应用新颖的方法来表征Gly- 罐，蛋白质及其结合相互作用。该提案中描述的研究很重要，因为它 将提供有关ESI-MS的基本观点，这项研究的分子见解将使 未来的ESI-MS方法的理性发展来表征这些分子。最终，这项工作将 促进细胞生物学和疾病状态中的聚糖，糖缀合物和蛋白质复合物的分析， 使这些方法能够应用于重要的生物学假设。