Protein Cross-Linking Technology and Applications

在化学系化学测量和成像项目的支持下，以及分子和细胞生物科学系细胞动力学和功能项目的部分共同资助下，印第安纳州大学的James Reilly教授和他的团队正在开发实验和计算方法，以更好地表征蛋白质之间的复杂相互作用，这些蛋白质对生物细胞的功能（和功能失调）至关重要。 他们的工作涉及相互作用，如蛋白质折叠和错误折叠（涉及阿尔茨海默病或帕金森病等疾病），以及与治疗某些疾病相关的抗体化学。 研究生，本科生和高中生谁参加这项研究了解物理，分析和生物化学的相互作用。他们还通过出版物和会议演讲参与传播研究成果，为他们在学术界、生物技术和制药业的进一步科学培训或职业生涯做好准备。交联质谱法是一种越来越受欢迎的研究蛋白质-蛋白质相互作用的方法。然而，蛋白质交联的分离和鉴定仍然充满挑战。赖利实验室的工作旨在通过提高蛋白质交联鉴定的置信度来为这一领域做出贡献。 多种碎片化方法，突出了新的交联剂，可裂解的电子转移解离，正在利用，以增加质谱数据中的信息内容。寻求定性和定量信息的数据融合是一个重大挑战。离子交换和等电聚焦用于富集和区分连接的蛋白质与未连接的对应物。蛋白质二聚热力学、蛋白质展开的温度依赖性以及核糖体与相关蛋白质之间的相互作用是测试硫代亚氨酸酯和市售琥珀酰亚胺酯交联剂的相对性能的潜在应用之一。该奖项反映了NSF的法定使命，并被认为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来支持。

With support from the Chemical Measurement and Imaging program in the Division of Chemistry, and partial co-funding from the Cellular Dynamics and Function program in the Division of Molecular and Cellular Biosciences, Professor James Reilly and his group at Indiana University are developing experimental and computational methods to better characterize the complex interactions among proteins that are critical to the function (and misfunction) of biological cells. Their work addresses interactions such as protein folding and misfolding (implicated in conditions such as Alzheimer's or Parkinson's disease), and the chemistry of antibodies relevant to treating certain diseases. Graduate students, undergrads and high school students who participate in this research learn about the interplay of physical, analytical and biochemistry. They are also involved in the dissemination of research results through publications and conference presentations that prepare them for further scientific training or careers in academia, biotechnology and the pharmaceutical industry.Cross-linking mass spectrometry is a method of studying protein-protein interactions that is growing in popularity. The isolation and identification of protein cross-links is nevertheless still filled with challenges. Work in the Reilly laboratory aims to contribute to this field by improving the degree of confidence of protein cross-link identifications. Multiple fragmentation methods, highlighted by novel cross-linkers that are cleavable by electron transfer dissociation, are being utilized to increase the information content in mass spectral data. Data fusion seeking both qualitative and quantitative information is a significant challenge. Ion exchange and isoelectric focusing are used to enrich and distinguish linked proteins from unlinked counterparts. Protein dimerization thermodynamics, the temperature dependence of protein unfolding, and interactions between ribosomes and associated proteins are among potential applications testing the relative performance of thioimidoesters and commercially available succinimidyl ester cross-linkers.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.