Deciphering Post-Translationally Modified Peptides with Arrayed Synthetic Receptors

在化学测量和成像（CMI）计划的支持下，以及化学系生命过程化学（CLP）和化学结构，动力学和机制-B（CSDM-B）计划的部分共同资助下，钟教授的团队和他们在加州大学的合作者-滨江正在开发和扩展表征蛋白质和肽合成后发生的化学修饰的方法。这种“翻译后修饰”（PTM）极大地扩展了蛋白质的结构和功能多样性。PTM之间的相互作用（“串扰”）对于蛋白质功能和生物过程的调节也是至关重要的。然而，PTM的鉴定和检测提出了重大挑战，因为多种类型和数量的PTM通常共存于单个肽或蛋白质上，并且这些不容易分离。一些修改只会对属性产生很小的变化，使得它们的检测变得困难。Zhong团队正在开发用于分析肽中重要类别PTM的创新工具，旨在增强我们对蛋白质修饰及其功能意义的理解。 该研究将与校园和外联活动相结合，旨在增加少数民族在STEM领域的参与，提高STEM领域服务不足的少数民族的保留率，并为对教育感兴趣的学生提供教学机会。 它将为研究生和本科生提供跨学科的研究培训，在化学，生物学和材料科学领域架起桥梁。Zhong团队正在开发与指示染料配对的灵活合成受体宿主，以研究PTM串扰并识别异构肽，这两项具有挑战性的任务目前的测量技术还不容易解决。采用多个正交识别机制，在具有上边缘和下边缘功能性的自折叠空腔与其荧光客体之间形成的阵列可以选择性地识别不同的PTM肽靶标。然后，开发的阵列可以用于研究PTM如何影响激酶的功能，负责将磷酸基团添加到蛋白质的酶，并研究PTM串扰，其中附近的修饰影响目标修饰的性质。此外，该阵列可用于检测异构肽之间的细微结构差异和测量异构酶的活性。通过以阵列形式利用多个不同的自折叠深腔体，这项工作旨在促进我们对PTM功能和调节的理解，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查进行评估，被认为值得支持的搜索.

With support from the Chemical Measurement and Imaging (CMI) program and partial co-funding from the Chemistry of Life Processes (CLP) and the Chemical Structures, Dynamics, and Mechanisms-B (CSDM-B) programs in the Division of Chemistry, Professor Zhong's group and their collaborators at the University of California-Riverside are developing and extending ways of characterizing chemical modifications occurring in proteins and peptides after they are synthesized. Such “post-translational modifications” (PTMs) greatly expand the structural and functional diversity of proteins. Interaction among PTMs (“crosstalk”) is also crucial for regulation of protein function and biological processes. However, identification and detection of PTMs pose significant challenges, because multiple types and numbers of PTMs often coexist on a single peptide or protein and these are not easily separable. Some modifications produce only small changes to the properties, making their sensing difficult. The Zhong group is developing innovative tools for the analysis of important classes of PTMs in peptides, aiming to enhance our understanding of protein modifications and their functional implications. The research will be integrated with on campus and outreach activities aiming to increase minority participation in STEM fields, to improve the retention rates of underserved minorities in STEM fields, and to provide teaching opportunities to students interested in education as a career goal. It will provide interdisciplinary research training to graduate students and undergraduates in areas that bridge chemistry, biology, and materials science.The Zhong group is developing flexible synthetic receptor hosts paired with indicator dyes to study PTM crosstalk and identify isomeric peptides, two challenging tasks not readily addressed by current measurement technologies. Employing multiple orthogonal recognition mechanisms, arrays formed between the self-folding cavitands with upper and lower rim functionality and their fluorescent guests can selectively recognize different PTM peptide targets. The developed arrays can then be employed to investigate how PTMs affect the function of kinases, enzymes responsible for adding phosphate groups to proteins, and to study PTM crosstalk, where nearby modifications influence the properties of a target modification. Additionally, the arrays can be used to detect the subtle structural differences among isomeric peptides and measure the activities of isomerases. By leveraging multiple different self-folding deep cavitands in an arrayed format, the work aims to advance our understanding of PTM functions and regulation, through improvements in PTM detection and by enabling the discovery of hosts with high affinity and selectivity for diverse PTM types.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.