多肽识别导向的线粒体及相关蛋白质靶向分析

Mitochondria have complicated and precise molecular interaction network and reaction systems. It is an important organelle where critical biological reactions take place. Targeted analysis based on the molecular features of mitochondria is crucial for the analysis and regulation of organelle functions, as well as detection of disease. In this proposal, mitochondria-related proteins and their molecular events are chosen as the targets. Peptides with high affinity and high selectivity will be designed and screened. Based on the peptide-protein recognition, novel materials will be developed for the rapid and specific separation of mitochondria and target proteins by using peptide as the affinity ligand. Guided by the targeting peptides, bioprobes will be designed and synthesized with responsive signal. Analytical methods for in-situ and real-time probing mitochondria and proteins in living cells can be developed. The peptide-based materials and bioprobes will be applied for monitoring the content, activity and distribution of mitochondria and target proteins in lesion cells or under stresses. The potential applications of the targeting peptide for detecting and interfering with mitochondrial activity will also be investigated. This study can provide promising materials and methods for detecting, understanding and regulating diseases at molecular level.

线粒体拥有一套复杂而精细的分子相互作用网络和分子反应系统，是细胞生命活动的重要场所。针对线粒体的分子特征开展靶向探测，对于细胞器功能解析、调控和疾病检测具有重要意义。本项目拟以线粒体特征蛋白质及其分子活动为靶标，开展高亲和力、高选择性靶向多肽的设计筛选。以多肽为界面识别分子，构建基于多肽-蛋白质相互作用的亲和分离材料，建立线粒体及靶标蛋白质快速、高选择性分离新方法。构建靶标分子识别响应的多肽生物探针，建立活细胞内线粒体和蛋白质的原位、动态传感分析新方法。利用分离材料和多肽探针，探测病变和应激状态下，蛋白质在细胞、线粒体水平的含量、活性和时空分布信息，探索靶向多肽在线粒体活性检测和调控中的新应用，为线粒体相关疾病检测、分子机制研究和干预提供新的思路与方法。

线粒体靶向分析和功能探测，对于认识线粒体复杂而精细的分子相互作用网络和分子反应系统具有重要意义，也是相关疾病检测和化学干预的关键基础。本项目研究以线粒体代谢和凋亡相关蛋白质为分子靶标，以蛋白-蛋白相互作用界面和功能结构域为靶点，设计合成了高亲和力、高选择性的小分子多肽。构建了遗传信息编码的多肽化合物库，建立了基于固-液异相和溶液均相的多肽亲和筛选新方法。以多肽为识别元件，设计构筑了配基多价定向功能化的微纳亲和界面和微阵列分离材料，建立了分子识别和形貌拓扑相互作用协同增效的线粒体分离新方法，实现了线粒体的选择性捕获和亚结构蛋白分析。利用线粒体特征负膜电位，构建了兼具亲脂和亲负电性的小分子及分子聚集体；将蛋白分子靶向、细胞器靶向和微环境靶向进行集成，设计合成了多靶向多肽生物探针，实现了复杂活体体系线粒体定位和功能蛋白分析。针对线粒体参与的细胞自噬、凋亡等关键生命过程，构建了结构可调、功能可控的新型靶向生物探针，建立了靶标蛋白识别响应和活性动态分析新方法，在活细胞和活体水平，实现了线粒体自噬、细胞凋亡等应激过程相关蛋白质表达水平、活性和分布变化的示踪分析，为疾病靶向探测、分子机制研究和化学干预提供新工具和新方法。本项目在Angew. Chem. Int. Ed., Anal. Chem.等学术期刊发表论文15篇，撰写英文专著1章，申请中国发明专利3项。项目执行期间，项目负责人获2021年国家自然科学基金优秀青年基金项目资助，获中科院化学所青年科学奖特别优秀奖2项，获中科院化学所青年科学奖优秀奖1项。