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Mechanisms and Applications of Protein Analysis by Desorption Electrospray Ionization-Mass Spectrometry (DESI-MS)

解吸电喷雾电离质谱 (DESI-MS) 蛋白质分析的机制和应用

基本信息

批准号：
2003379
负责人：
Andre Venter
金额：
$ 39.68万
依托单位：
Western Michigan University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2003379&HistoricalAwards=false
关键词：
Mechanisms Applications Protein Analysis Desorption

项目摘要

With support from the Chemical Measurement and Imaging program in the Division of Chemistry, Professors Andre Venter and Todd Barkman’s team at Western Michigan University are devising sensitive methods for identifying and characterizing proteins - key building blocks of life. By systematically optimizing sample additives and probe parameters for analysis of proteins in solution and on surfaces, Dr. Venter and his team is not only improving protein analysis, but is also adding to the fundamental understanding of protein solubility and stability, with important potential consequences for bioanalytical measurements, recombinant protein expression, and applications such as the development of protein therapeutics. Students participating in these studies learn about chemistry and biology. In addition, Dr. Venter and colleagues bring the research topics into the classroom, including an unusual interdisciplinary course engaging aspiring scientists with students in the sonic and visual arts. These exposures help prepare both groups for the interdisciplinary nature of STEAM - science, technology, engineering, arts and mathematics - in contemporary society. This research draws on the observation that adding serine either to the spray or to a deposited sample can enhance sensitivity in desorption electrospray ionization (DESI) mass spectrometric analysis of proteins. The underlying mechanism is assessed by studying the effects of derivatization of the serine additive at all three reactive sites. Signals derived from protein monomers and aggregates in native and denatured states are monitored under basic, neutral, and acidic conditions to probe which functional groups and molecular properties drive protein dissolution and stability in solution. Drs. Venter and Barkman are also studying the efficacious effect of adding ethyl acetate vapor to the atmosphere surrounding the analyte, which has been found to increase protein signal intensities and reduce the abundance of spectral features derived from clusters. Better understanding of the mechanisms of these effects may enable novel high-throughput strategies for identification of proteins and their enzymatic functions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系化学测量和成像项目的支持下，西密歇根大学的安德烈·文特尔教授和托德·巴克曼团队正在设计识别和表征蛋白质的灵敏方法--蛋白质是生命的关键组成部分。通过系统地优化样品添加剂和探针参数，用于分析溶液中和表面上的蛋白质，Venter博士和他的团队不仅改进了蛋白质分析，而且还增加了对蛋白质溶解度和稳定性的基本理解，对生物分析测量，重组蛋白表达和应用（如蛋白质治疗剂的开发）具有重要的潜在影响。参加这些研究的学生学习化学和生物学。此外，文特尔博士及其同事将研究课题带入课堂，包括一门不同寻常的跨学科课程，让有抱负的科学家与声音和视觉艺术的学生一起学习。这些接触有助于两组准备蒸汽的跨学科性质-科学，技术，工程，艺术和数学-在当代社会。这项研究借鉴的观察，添加丝氨酸的喷雾或沉积的样品可以提高灵敏度的解吸电喷雾电离（DESI）质谱分析的蛋白质。通过研究丝氨酸添加剂在所有三个反应位点的衍生化的影响来评估潜在的机制。在碱性、中性和酸性条件下监测来自天然和变性状态的蛋白单体和聚集体的信号，以探测哪些官能团和分子特性驱动蛋白在溶液中的溶解和稳定性。Venter和Barkman博士还在研究向分析物周围的大气中加入乙酸乙酯蒸气的有效效果，发现这会增加蛋白质信号强度并减少来自簇的光谱特征的丰度。更好地了解这些影响的机制可能使新的高通量策略，用于识别蛋白质及其酶功能。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。