自噬检测指南讲述.pptx

细胞自噬研究方法概述 研究生：王颖 导师：刘乃丰 教授 Autophagic Compartments Autophagic Compartments Phagophore (pre-autophagosomal)： previously called the isolation or sequestration membrane 吞噬泡 ：参与自噬体形成早期事件的膜池。也指“隔离膜isolation membrane”或“杯状结构cup-shaped structure” Autophagosome : 自噬体 ：双层膜包裹胞质形成的囊泡 Amphisome ：generated by the fusion of autophagosomes with endosomes, also referred to as an acidic late autophagosome 自噬内涵体：溶酶体和内涵体融合的中间囊泡 Autolysosome ：generated by fusion of autophagosomes or amphisomes with a lysosome 自噬溶酶体：自噬小体和溶酶体融合形成的终末结构 Autophagic molecular mechanisms Initiation： Induction，Cargo recognition and selectivity Elongation，Closure： Autophagosome formation Maturation，Degradation: Vesicle fusion and autophagosome breakdown Autophagic molecular mechanisms Induction Normal conditions Basal-level autophagy is very low; Autophagy inhibitor: serine/threonine protein kinase TOR(target of rapamycin) input information from multiple upstream signal transduction pathways (discussed below) and negatively regulates another serine/threonine kinase, Atg1, in nutrient-rich conditions Starvation conditons or Rapamycin TOR inhibited; Atg1 activated; Atg1 binding affinity to Atg13 and Atg17 ↑; Promotes the formation of an Atg1-Atg13-Atg17 scaffold ; Atg1-Atg13-Atg17 recruitment of multiple Atg proteins to the PAS to initiate autophagosome formation. Cargo recognition and selectivity P62/sequestosome 1 (SQSTM1) . P62 directly binds both poly- or mono-ubiquitin via its ubiquitin-associated (UBA) domain and LC3 links the ubiquitinated cargos to the autophagy machinery for autophagic degradation.  Class III phosphatidylinositol 3-kinase (PtdIns3K) complex: PtdIns3K Vps34 (vacuolar protein sorting 34), a myristoylated serine/threonine kinase Vps15, Atg14 ; Beclin 1 ; Autophagosome formation  Vps34 Atg14 Vps15 Beclin1 The PtdIns3K complex produces PtdIns3P (phosphatidylinositol 3-phosphate) and is involved in PAS targeting of a number of yeast Atg proteins that bind PtdIns3P, s