Ultraviolet Photodissociation Mass Spectrometry for Characterization of Biological Molecules

用于表征生物分子的紫外光解离质谱法

• 批准号: 10797256
• 负责人: Jennifer S. Brodbelt
• 金额: $ 16万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797256
• 关键词: Antibiotic Resistance; Area; Binding; Biological; Biomedical Research; C-terminal; Capillary Electrophoresis; Cellular Membrane; Chemicals; Collaborations; Complex; Disease; Funding; Genetic Transcription; Goals; Hybrids; Individual; Ligands; Lipids; Mass Spectrum Analysis; Membrane Proteins; Methods; Microbiology; Modification; Molecular; Molecular Biology; Nucleic Acids; Pattern; Phosphorylation; Post-Translational Protein Processing; Proteins; RNA Polymerase II; Research; Structure; Technology; cost; design; frontier; functional outcomes; innovation; insight; macromolecular assembly; mass spectrometer; novel therapeutics; oxidation; pathogenic bacteria; programs; protein complex; protein function; tandem mass spectrometry; ultraviolet

Abstract. Understanding the functions of lipids, proteins and even larger macromolecular assemblies depends on deciphering complex structures of individual molecules as well as decrypting how those molecules interact, often via networks of non-covalent interactions. In order to advance the elucidation of biomolecular organization and functional outcomes, new methods are needed to push the limits of structural insight, providing more detailed holistic chemical information with greater sensitivity. The critical interplay between structure/function is evidenced in numerous biologically-motivated problems, ranging from understanding the ways that pathogenic bacteria develop antibiotic resistance to the design of new drugs that selectively bind and inhibit the functions of protein targets. The ongoing need for even greater chemical insight has motivated my group’s effort to develop innovative mass spectrometry methods to characterize structures of biological molecules in unprecedented detail, especially lipids and proteins which are featured in this proposal. The overarching goal of my research program is to develop state-of- the-art tandem mass spectrometry technologies, particularly highlighting ultraviolet photodissociation (UVPD) and hybrid MS/MS methods, for structural elucidation of lipids, proteins, and protein complexes. These new methods will be showcased for solving challenging problems in three areas. (1) Lipids: (i) profiling lipids of pathogenic bacteria and their signatures of antibiotic resistance, and (ii) structural characterization of unsaturations, oxidations and modifications of lipids that occur during remodeling of cellular membranes. (2) Protein complexes: (i) characterization of protein-ligand complexes, membrane protein complexes, protein/nucleic acid complexes, and macromolecular assemblies, and (ii) advancing capillary electrophoresis for native separations and exploration of the interactome. (3) Post-translational modifications: focusing on decoding the phosphorylation patterns of the C-terminal domain of RNA polymerase II which regulates transcription. These high impact problems are supported via numerous collaborations with microbiology and molecular biology groups who recognize the value of frontier mass spectrometry strategies for elevating biomedical research. This supplement supports acquisition of a Thermo QE-HFX Orbitrap mass spectrometer that will be used for all three project areas and for which the cost will be heavily leveraged by other funds.

抽象的。了解脂质、蛋白质甚至更大的大分子组装体的功能 取决于破译单个分子的复杂结构，以及这些分子是如何 分子通常通过非共价相互作用的网络相互作用。为了进一步阐明 生物分子组织和功能结果，需要新的方法来推动限制， 结构洞察力，提供更详细的整体化学信息，具有更高的灵敏度。临界 结构/功能之间的相互作用在许多生物学动机的问题中得到证明， 从了解致病菌产生抗生素耐药性的方式，到设计新的 选择性结合并抑制蛋白质靶点功能的药物。持续需要更大的 化学洞察力促使我的团队努力开发创新的质谱分析方法， 以前所未有的细节描述生物分子的结构，特别是脂质和蛋白质 这是本提案中的特色。我的研究计划的首要目标是发展国家的- 最先进的串联质谱技术，特别强调紫外光解离 （UVPD）和混合MS/MS方法，用于脂质、蛋白质和蛋白质复合物的结构解析。 这些新方法将被展示用于解决三个领域的挑战性问题。(1)脂质：（i） 分析病原菌的脂质及其抗生素抗性的特征，和（ii）结构 表征重塑过程中发生的脂质的不饱和、氧化和修饰， 细胞膜(2)蛋白质复合物：（i）蛋白质-配体复合物的表征，膜 蛋白质复合物、蛋白质/核酸复合物和大分子组装体，以及（ii）推进 毛细管电泳用于天然分离和相互作用组的探索。(3)翻译后 修饰：专注于解码RNA C末端结构域的磷酸化模式 调节转录的聚合酶II。这些高影响力的问题得到了许多支持， 与认识到前沿质量价值的微生物学和分子生物学小组合作 光谱分析战略，以提升生物医学研究。此补充支持购买 Thermo QE-HFX轨道阱质谱仪将用于所有三个项目区域， 其他基金将大量利用这笔费用。

项目成果

Structural Characterization and Quantitation of Ether-Linked Glycerophospholipids in Peroxisome Biogenesis Disorder Tissue by Ultraviolet Photodissociation Mass Spectrometry.

• DOI: 10.1021/acs.analchem.2c01274
• 发表时间: 2022-09-20
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Blevins, Molly S.;Shields, Samuel W. J.;Cui, Wei;Fallatah, Wedad;Moser, Ann B.;Braverman, Nancy E.;Brodbelt, Jennifer S.
• 通讯作者: Brodbelt, Jennifer S.

Enhancing the Signal-to-Noise of Diagnostic Fragment Ions of Unsaturated Glycerophospholipids via Precursor Exclusion Ultraviolet Photodissociation Mass Spectrometry (PEx-UVPD-MS).

• DOI: 10.1021/acs.analchem.2c02128
• 发表时间: 2022-08-16
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Shields, Samuel W. J.;Sanders, James D.;Brodbelt, Jennifer S.
• 通讯作者: Brodbelt, Jennifer S.

Deciphering combinatorial post-translational modifications by top-down mass spectrometry.

• DOI: 10.1016/j.cbpa.2022.102180
• 发表时间: 2022-10
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8

Sialidase NEU3 action on GM1 ganglioside is neuroprotective in GM1 gangliosidosis.

Sialidase NEU3对GM1神经苷的作用在GM1神经节病中具有神经保护作用。

• DOI: 10.1016/j.jlr.2023.100463
• 发表时间: 2023-12
• 期刊: JOURNAL OF LIPID RESEARCH
• 影响因子: 6.5
• 作者: Allende, Maria L.;Lee, Y. Terry;Byrnes, Colleen;Li, Cuiling;Tuymetova, Galina;Bakir, Jenna Y.;Nicoli, Elena-Raluca;James, Virginia K.;Brodbelt, Jennifer S.;Tifft, Cynthia J.;Proia, Richard L.
• 通讯作者: Proia, Richard L.

Ultraviolet Photodissociation Permits Comprehensive Characterization of O-Glycopeptides Cleaved with O-Glycoprotease IMPa.

紫外线光解可以对 O-糖蛋白酶 IMPa 切割的 O-糖肽进行全面表征。

• DOI: 10.1021/acs.analchem.3c01111
• 发表时间: 2023
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Helms,Amanda;Escobar,EdwinE;Vainauskas,Saulius;Taron,ChristopherH;Brodbelt,JenniferS
• 通讯作者: Brodbelt,JenniferS