权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Novel Methods to Study Metastable Biomolecular Systems with Native LC/MS

使用天然 LC/MS 研究亚稳态生物分子系统的新方法

基本信息

批准号：
1709552
负责人：
Igor Kaltashov
金额：
$ 42万
依托单位：
University of Massachusetts Amherst
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2021-05-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1709552&HistoricalAwards=false
关键词：
Novel Methods Study Metastable Biomolecular

项目摘要

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Prof. Igor Kaltashov and his group at the University of Massachusetts at Amherst are developing powerful analytical tools to study processes such as protein/receptor recognition and enzymatic reactions. These new tools will advance understanding of the mechanisms of complex biological processes at the molecular level, ultimately facilitating manipulation of these processes to achieve desired outcomes. Thus, the new tools can have significant impact not only in chemistry, but also in biotechnology and medicine. Elements of the research involve close interaction with industrial collaborators, adding a valuable dimension to the educational experience of the graduate and undergraduate students involved. Dr. Kaltashov and his group are also engaging in outreach activities which seek to expose local high school students to the state-of-the-art analytical instrumentation at UMass-Amherst.Characterization of biopolymer dynamics and architecture (conformations of individual biopolymer chains and assemblies of multi-unit systems) is both challenging and important for improved understanding of biochemical processes. There is particular need for tools capable of probing transient systems, which are encountered in processes such as protein aggregation, assembly and evolution of quaternary structures, interaction with physiological partners, and enzyme catalysis and transport phenomena. Improved understanding of these phenomena requires reliable kinetics data and knowledge of conformational changes that accompany transitions. In order to improve on existing characterization capabilities, the Kaltashov group is developing a new robust, sensitive, and highly selective experimental strategy that uses a combination of liquid chromatography (LC) run under native conditions (e.g., size-exclusion chromatography) with on-line detection by native electrospray ionization mass spectrometry (MS). The new LC/MS platform incorporates ion manipulation in the gas phase to enable studies of highly heterogeneous systems (e.g., protein-polymer conjugates). Incorporation of on-column H/D exchange and chemical reduction reactions enables top-down HDX MS/MS characterization of higher order structure and conformational dynamics of proteins.

在化学测量和成像项目的支持下，马萨诸塞大学阿默斯特分校的Igor Kaltashov教授和他的团队正在开发强大的分析工具来研究蛋白质/受体识别和酶促反应等过程。这些新工具将在分子水平上推进对复杂生物过程机制的理解，最终促进对这些过程的操纵，以实现预期的结果。因此，这些新工具不仅在化学领域，而且在生物技术和医学领域都能产生重大影响。研究的要素包括与工业合作者的密切互动，为所涉及的研究生和本科生的教育经验增加了宝贵的维度。Kaltashov博士和他的团队还参与了一些拓展活动，旨在让当地的高中生接触到马萨诸塞大学阿默斯特分校最先进的分析仪器。生物聚合物动力学和结构（单个生物聚合物链的构象和多单元系统的组装）的表征对提高对生化过程的理解既具有挑战性又很重要。特别需要能够探测瞬时系统的工具，这些系统在蛋白质聚集、四级结构的组装和进化、与生理伙伴的相互作用以及酶催化和运输现象等过程中都会遇到。提高对这些现象的理解需要可靠的动力学数据和伴随转变的构象变化的知识。为了提高现有的表征能力，Kaltashov小组正在开发一种新的强大、敏感和高选择性的实验策略，该策略使用液相色谱（LC）在天然条件下运行（例如，尺寸排除色谱）与天然电喷雾电离质谱（MS）在线检测相结合。新的LC/MS平台结合了气相离子操作，可以研究高度非均相系统（例如，蛋白质-聚合物偶联物）。结合柱上H/D交换和化学还原反应，可以自上而下的HDX MS/MS表征蛋白质的高阶结构和构象动力学。