Laboratorio de Biología Celular, Centro de Investigación Príncipe Felipe, Valencia, Spain.
J Cell Sci. 2012 Jan 1;125(Pt 1):92-107. doi: 10.1242/jcs.086728. Epub 2012 Jan 20.
Macroautophagy is a major lysosomal catabolic process activated particularly under starvation in eukaryotic cells. A new organelle, the autophagosome, engulfs cytoplasmic substrates, which are degraded after fusion with endosomes and/or lysosomes. During a shotgun proteome analysis of purified lysosomal membranes from mouse fibroblasts, a Ca(2+)-dependent phospholipid-binding protein, annexin A5, was found to increase on lysosomal membranes under starvation. This suggests a role for this protein, an abundant annexin with a still unknown intracellular function, in starvation-induced lysosomal degradation. Transient overexpression and silencing experiments showed that annexin A5 increased lysosomal protein degradation, and colocalisation experiments, based on GFP sensitivity to lysosomal acidic pH, indicated that this was mainly the result of inducing autophagosome-lysosome fusion. Annexin A5 also inhibited the endocytosis of a fluid-phase marker and cholera toxin, but not receptor-mediated endocytosis. Therefore, we propose a double and opposite role of annexin A5 in regulating the endocytic and autophagic pathways and the fusion of autophagosomes with lysosomes and endosomes.
巨自噬是真核细胞在饥饿时特别激活的主要溶酶体分解代谢过程。一种新的细胞器,自噬体,吞噬细胞质底物,这些底物在与内体和/或溶酶体融合后被降解。在对从小鼠成纤维细胞纯化的溶酶体膜进行的鸟枪法蛋白质组分析中,发现钙依赖性磷脂结合蛋白 annexin A5 在饥饿时在溶酶体膜上增加。这表明该蛋白(一种丰富的 annexin,其细胞内功能仍未知)在饥饿诱导的溶酶体降解中发挥作用。瞬时过表达和沉默实验表明 annexin A5 增加了溶酶体蛋白降解,并且基于 GFP 对溶酶体酸性 pH 的敏感性的共定位实验表明,这主要是由于诱导自噬体-溶酶体融合所致。Annexin A5 还抑制了液相间质标记物和霍乱毒素的内吞作用,但不抑制受体介导的内吞作用。因此,我们提出 annexin A5 在调节内吞作用和自噬途径以及自噬体与溶酶体和内体融合方面具有双重相反的作用。