CAREER: Towards an Understanding of the Underlying Chemistry in Collision-Induced Dissociation of Peptides in Mass Spectrometry

来自佛罗里达大学（UF）的Nicolas Polfer得到化学系分析和表面化学项目的支持，研究肽的碰撞诱导解离（CID），这是肽和蛋白质质谱测序和鉴定的基石。 在CID的高能量条件下，一些片段可以经历重排过程，使得原始序列信息丢失。 这种“混乱”过程被认为是通过N-末端的环化反应进行的。 Polfer博士的研究小组采用红外光解离光谱法来鉴定形成的碎片结构，并采用离子迁移率质谱法来分离和定量CID产物的通常不均匀的混合物。 这些研究试图确定序列“扰乱”和/或其他过程是否普遍存在，从而引起对肽错误识别的关注。该项目使佛罗里达大学的本科生和佛罗里达的高中教师接触生命科学和生命科学技术（如质谱）通过霍华德休斯医学研究所的生命科学计划，在那里，本科生被介绍给高度跨学科的方法和技术，对生物技术很重要。 PI还正在开发模块，重点关注与蛋白质鉴定和质谱法相关的尖端技术的应用，用于UF大学前教育和培训中心组织的夏季教师研讨会。 鼓励教师将获得的知识和技能转移到他们的高中课堂;一个后续的春季讲习班遵循他们的进展。 该项目支持来自贫困地区的教师参与。

Nicolas Polfer from the University of Florida (UF) is supported by the Analytical and Surface Chemistry Program in the Chemistry Division to investigate the collision-induced dissociation (CID) of peptides, a cornerstone in mass spectrometric sequencing and identification of peptides and proteins. Under the high-energy conditions of CID, some fragments can undergo rearrangement processes, such that the original sequence information is lost. This "scrambling" process is thought to operate via a cyclization reaction from the N-terminus. Dr. Polfer's group employs infrared photodissociation spectroscopy to identify the fragment structures formed, and ion mobility mass spectrometry to separate and quantify the often heterogeneous mixtures of CID products. The studies seek to determine whether sequence "scrambling" and/or other processes are pervasive and thus a concern for peptide misidentification.The project exposes undergraduate students at UF and high-school teachers in Florida to life sciences and life science-enabling technologies (such as mass spectrometry) via the Howard Hughes Medical Institute Science for Life program, where undergraduates are introduced to highly interdisciplinary approaches and techniques important for biotechnology. The PI is also developing modules focused on the application of cutting-edge technologies related to protein identification and mass spectrometry for summer teacher workshops organized by the Center for Precollegiate Education and Training at UF. Teachers are encouraged to transfer derived knowledge and skills to their high-school classrooms; a follow-up spring workshop follows their progress. The project supports participation by teachers from disadvantaged areas.