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

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·巴克曼教授的团队正在设计敏感的方法来识别和表征蛋白质--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以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，还增加了对蛋白质溶解性和稳定性的基本了解，对生物分析测量、重组蛋白质表达和蛋白质疗法开发等应用具有重要的潜在影响。参与这些研究的学生学习化学和生物。此外，文特尔博士和他的同事们把研究课题带到了课堂上，包括一门不同寻常的跨学科课程，让有抱负的科学家与声学和视觉艺术的学生互动。这些经历有助于这两个群体为当代社会蒸汽的跨学科性质--科学、技术、工程、艺术和数学--做好准备。在蛋白质的解吸电喷雾电离(DESI)质谱分析中，将丝氨酸添加到喷雾或沉积样品中可以提高灵敏度。通过研究丝氨酸添加剂在所有三个反应部位的衍生化效果来评估潜在的机理。在碱性、中性和酸性条件下，监测来自天然和变性状态下的蛋白质单体和聚集体的信号，以探索哪些官能团和分子性质推动蛋白质在溶液中的溶解和稳定性。文特尔和巴克曼博士还在研究向分析物周围的大气中添加乙酸乙酯蒸汽的有效效果，已发现这会增加蛋白质信号强度，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会使鉴定蛋白质及其酶功能的新的高通量策略成为可能。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.