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

天然质谱学指导结构生物学P41资源(NMS-&GT；SB资源)， 成立于2018年，是一个国家生物医学技术研究资源，旨在开发和 传播结构生物学的新研究技术和方法。该资源正在开发和使用 在五个技术研究与开发项目下传播一套技术；这些 包括(R&D 1-3)耦合到高分辨率离子迁移率(IM)新型表面诱导解离(SID)单元 和高分辨质谱学(HRMS)、在线纯化和分离方案(R&D 4)，以及 (R&D 5)用于从实验NMS数据确定结构的计算工具。该资源具有 成功地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：蛋白质复合体，包括合成的淀粉样β蛋白 (aβ)和tau聚集体，来自贝克实验室的全新设计的蛋白质复合体(加州大学 华盛顿)，由Sharon实验室(魏兹曼研究所)表达的同源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)。我们的蛋白质：蛋白质和 核蛋白复合体通常以对质谱学友好的浓度和质量运行 光谱兼容的缓冲液/电解液。仪器的获取(Refeyn OneMP质谱仪) 对于基于光的质量光度测量(干涉测量)将为我们的DBP和 C&S调查人员，使我们能够在非常低的浓度(PM到低NM)下评估小样本量。质量 光度学(MP)将提供样品质量和样品中低聚物分布的快速筛选 在调查员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 为了样品。我们将决定是将样本送回调查人员进行改进，还是 是否为更高分辨率、更高质量精度的原生质谱学测量做好准备。为. 多种多样的蛋白质复合体类型，由DBP和C&S项目提供，我们将建立 MP和NMS结果重复的复杂类型和集中行为 这两组结果提供了相加/互补的数据，以在 大范围的条件(齐聚时间/动力学、浓度、缓冲条件)。

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).