CAREER: A Chemical Technology to Define Target-Specific Bioreactivity: Integrating Research and Education at the Crossroads of Chemistry and Biology

细胞内部的工作需要对一系列单独的化学事件进行精细的协调。各种各样的“仪器”通过无数的信号和分子相互交流，这些信号和分子传递着细胞状态的信息。对于细胞的正常功能来说，信号的时间决定一切。为了了解活细胞内部发生了什么，研究人员经常将信号分子应用于细胞并观察发生了什么。这种方法有严重的局限性，因为整个细胞都暴露在信号中，而且随着化合物进入细胞内部，时间在很大程度上丢失了。这些影响可能导致意想不到的后果，而这些后果可能是无法解决的。该提案旨在克服这些限制，并获得对特定信号分子释放的确切位置和时间的控制。研究人员通过开发一套新的化学工具来解决这一迫切需求，这些化学工具可以在精确的时间选择性地将反应信号传递给细胞中的特定蛋白质。这一研究突破将为生物学中具有重要基础意义的化学定向信号处理机制提供时间分辨信息。这项研究使实验室的研究生能够在化学和生物学的技术开发方面获得良好的基础。综合推广目标是突出现代横切化学生物学概念和工具，特别针对K-12教育工作者和来自代表性不足人群的学生。化学部的生命过程化学项目正在资助康奈尔大学的助理教授Yimon Aye，以时空精度评估基本的蛋白质/小分子信号相互作用。由生物活性信号亲电试剂（如脂质衍生醛）诱导的非酶翻译后蛋白修饰在氧化还原连接的细胞信号传导中起着重要作用。为了避免传统的全细胞治疗方法，本提案引入了一种新的化学驱动方法，通过在一个天然/健康细胞中，在精确的时间对一个靶标和一个诱导剂进行选择性的化学扰动，来询问这些相互作用。来自PI实验室的最新数据表明，她的新定向修饰策略可以定量评估活细胞中单个非酶辅助蛋白质修饰事件的靶和亲电特异性时间动态。生理相关反应性和特异性的知识将使预测一个给定的反应性小分子信号如何在细胞决策目标的微环境中发挥作用。

The inner workings of cells require exquisite orchestration of an array of individual chemical events. The various "instruments" communicate with one another through numerous signals, molecules that convey information about the state of the cell. For the proper function of cells, the timing of signaling is everything. In order to understand what is going on inside live cells, researchers routinely apply the signaling molecules to cells and observe what happens. There are serious limitations to this approach because the entire cell is exposed to the signal and timing is largely lost as the compounds find their way inside. These effects can lead to unintended consequences that can be impossible to sort out. The proposal aims to overcome these limitations, and to gain control over the exact location and timing of release of specific signaling molecules. The investigator addresses this pressing need by developing a set of new chemical tools with which reactive signals can be selectively delivered to specific proteins in cells at a precise time. Such a research breakthrough will provide time-resolved information on the chemically directed signal-processing mechanisms of fundamental importance in biology. The research enables graduate students in the lab to acquire excellent grounding in technological development interfacing chemistry and biology. The integrated outreach goal is to highlight modern crosscutting chemical biology concepts and tools, specifically aimed at K-12 educators and students from underrepresented populations.With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Assistant Professor Yimon Aye from Cornell University to evaluate with spatiotemporal precision the fundamental protein/small-molecule signal interactions. The non-enzymatic post-translational protein modifications inducible by the bioactive signaling electrophiles, such as lipid-derived aldehydes, have emerged to play major roles in redox-linked cell signaling. Avoiding the conventional whole-cell treatment approaches, this proposal introduces a new chemistry-driven approach to interrogate these interactions by selective chemical perturbation of one target with one inducer at a precise time in an otherwise native/healthy cell. The most recent data from the PI's lab suggest that her new directed modification strategy enables quantitative assessment of target- and electrophile-specific temporal dynamics of individual non-enzyme-assisted protein modification events in living cells. The knowledge of physiologically relevant reactivity and specificity will enable the prediction of how a given reactive small-molecule signal may function within microenvironments of a target in cellular decision-making.