Resource for native mass spectrometry guided structural biology-Refeyn OneMP Mass Photometer

原生质谱引导结构生物学资源 -Refeyn OneMP 质量光度计

• 批准号: 10400442
• 负责人: Vicki H. Wysocki
• 金额: $ 13.6万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400442
• 关键词: Alzheimer' s Disease; Amyloid beta-Protein; Automobile Driving; Back; Behavior; Biomedical Technology; Buffers; Cells; Chemicals; Collaborations; Complement; Complex; Core Facility; Country; Coupled; Cryoelectron Microscopy; DNA; Data; Devices; Dissociation; Ebola virus; Electrolytes; Equipment; Heterogeneity; Imaging Device; Immunology; Institutes; Interferometry; Kinetics; Laboratories; Light; Mass Spectrum Analysis; Measurement; Measures; Methodology; Methods; Mission; Modeling; Molecular Chaperones; Molecular Conformation; Monitor; Nucleoproteins; Organism; Peptides; Performance; Photometry; Polymorph; Protein Engineering; Proteins; RNA; RNase P; Rapid screening; Research; Research Personnel; Resolution; Resources; Running; Sampling; Scheme; Services; Structure; Surface; System; Technology; Time; Training; United States National Institutes of Health; Universities; Washington; Work; aggregation pathway; base; computerized tools; equipment acquisition; instrument; ion mobility; member; multicatalytic endopeptidase complex; nanodisk; new technology; novel; protein complex; rho; single molecule; structural biology; tandem mass spectrometry; tau Proteins; tau aggregation; technology research and development; tool

The P41 Resource for Native Mass Spectrometry Guided Structural Biology (nMS->SB Resource), established in 2018, is a national Biomedical Technology Research Resource that seeks to develop and disseminate novel research technologies and methods for structural biology. The Resource is developing and disseminating a suite of technologies under five Technology Research and Development (TR&D) projects; these include (TR&D 1-3) novel surface-induced dissociation (SID) cells coupled to high-resolution ion mobility (IM) and high-resolution mass spectrometry (HRMS), (TR&D 4) online purification and separation schemes, and (TR&D 5) computational tools for structure determination from experimental nMS data. The Resource has successfully supported the efforts of numerous biomedical researchers across the country and the globe to characterize a variety of noncovalent protein and RNA/DNA:protein complexes, including synthetic amyloid beta (Aβ) and tau aggregates, novel de novo designed protein complexes from the Baker laboratory (University of Washington), orthologous 20S proteasomes expressed by the Sharon laboratory (Weizmann Institute) and recently Ebola virus matrix protein VP40 (La Jolla Institute of Immunology). Our protein:protein and nucleoprotein complexes are typically run at mass spectrometry-friendly concentrations and in mass spectrometry-compatible buffers/ electrolytes. Acquisition of an instrument (Refeyn OneMP Mass Photometer) for light-based mass photometry (interferometry) measurements will provide new technology for our DBP and C&S investigators, allowing us to assess small sample volumes at very low concentrations (pM to low nM). Mass photometry (MP) will provide a quick screen of sample quality and oligomeric distributions present in the sample in the investigator’s provided storage buffer, allowing MS team members to decide on the appropriate next steps for the sample. We will determine whether the sample will go back to the investigator for improvements or whether it is ready for higher resolution, higher mass accuracy native mass spectrometry measurements. For a wide variety of protein complex types, provided by a broad set of DBP and C&S projects, we will establish complex types and concentration behaviors for which the MP and nMS results are duplicative and those for which the two sets of results provide additive/complementary data to characterize the complex more fully over a large range of conditions (oligomerization time/kinetics, concentrations, buffer conditions).

天然质谱引导结构生物学的 P41 资源（nMS->SB 资源）， 成立于2018年，是国家生物医学技术研究资源中心，致力于开发和 传播结构生物学的新颖研究技术和方法。 在五个技术研究与开发（TR&D）项目下传播一套技术； 包括（TR&D 1-3）与高分辨率离子淌度（IM）耦合的新型表面诱导解离（SID）细胞 和高分辨率质谱 (HRMS)、(TR&D 4) 在线纯化和分离，以及 (TR&D 5) 用于根据实验 nMS 数据确定结构的计算工具。 成功支持了全国和全球众多生物医学研究人员的努力 表征各种非共价蛋白质和 RNA/DNA:蛋白质复合物，包括合成的淀粉样蛋白 beta (Aβ) 和 tau 聚集体，贝克实验室（University of Baker）的新型从头设计的蛋白质复合物 Washington），Sharon 实验室（魏茨曼研究所）表达的直系同源 20S 蛋白酶体和 最近埃博拉病毒基质蛋白VP40（拉霍亚免疫学研究所）：蛋白质和。 核蛋白复合物通常在适合质谱分析的浓度和质量下运行 购买仪器（Refeyn OneMP 质量光度计）。 基于光的质量光度测量（干涉测量）将为我们的 DBP 和 C&S 研究人员，使我们能够评估极低浓度（pM 至低 nM）的小样本量。 光度测定 (MP) 将快速筛选样品质量和样品中存在的低聚物分布 在研究者提供的存储缓冲区中，允许 MS 团队成员决定适当的后续步骤 我们将决定是否将样本返回给研究者进行改进或 是否已准备好进行更高分辨率、更高质量精度的原生质谱测量。 由一系列广泛的 DBP 和 C&S 项目提供的各种蛋白质复合物类型，我们将建立 MP 和 nMS 结果重复的复杂类型和浓度行为以及 这两组结果提供了附加/互补数据，可以更全面地描述复杂的特征 大范围的条件（寡聚时间/动力学、浓度、缓冲液条件）。