目前，基于细胞自噬小体特异性标记蛋白LC3-PE，显微成像研究自噬小体与其他细胞器互作网络的方法有两种：LC3抗体免疫荧光标记和GFP-LC3融合蛋白。LC3抗体能同时识别LC3-I 和LC3-PE，显微镜下不能区分胞浆GFP-LC3和包含GFP-LC3-PE的微小囊泡。本项目拟开发多种新型的特异性检测脂基化蛋白LC3-PE的化学生物学工具，用于研究细胞自噬小体与其他细胞器互作网络。项目拟制备LC3-PE的特异性抗体；结合免疫荧光技术，显微成像研究细胞自噬小体与其他细胞器的互作；拟合成包含炔基的磷脂酰乙醇胺（alkyne-PE）探针，利用代谢标记、生物正交反应和分子内FRET技术，特异性检测LC3-PE，从而动态检测细胞自噬小体与其他细胞器互作。项目预期将开发新型的检测LC3-PE和细胞自噬小体的化学生物学工具，项目结果将揭示细胞器互作对细胞自噬小体的形成和细胞自噬进程的调节机制。

Currently, based on the autophagy-specific marker protein LC3-PE, there are two methods for the study of the inter-organelle communication of autophagosome and other organelles: LC3 antibody immunofluorescence labeling and GFP-LC3 fusion protein. The LC3 antibody recognizes both LC3-I and LC3-PE, and the cytoplasmic GFP-LC3 and vesicles containing GFP-LC3-PE cannot be distinguished under the microscope. The aim of this project is to develop novel chemical biology tools for the specific detection of lipidated protein LC3-PE for the study of the inter-organelle communication of autophagosomes and other organelles. Firstly, the project intends to prepare specific antibodies for LC3-PE. In combination with immunofluorescence techniques, the inter-organelle communication between autophagosomes and other organelles will be addressed. Secondly, the chemical probe alkyne-PE will be synthesized. Using metabolic labelling, bio-orthogonal reactions, and intramolecular FRET technology, LC3-PE will be specific detected and further dynamic interactions between autophagosomes and other organelles will be explained. The project is expected to develop a new chemical biology tools for the detection of LC3-PE and autophagosomes. The results of the project will reveal the role of inter-organelle communication in the formation of autophagosome and during autophagy processes.