Molecular Membrane Biology Group, Department of Biology/Chemistry, Osnabrück University, Osnabrück, Germany.
Institute of Cell Dynamics and Imaging, University of Münster, Münster, Germany.
J Cell Biol. 2022 Apr 4;221(4). doi: 10.1083/jcb.202107148. Epub 2022 Feb 17.
The lysosome is the major catabolic organelle in the cell that has been established as a key metabolic signaling center. Mutations in many lysosomal proteins have catastrophic effects and cause neurodegeneration, cancer, and age-related diseases. The vacuole is the lysosomal analog of Saccharomyces cerevisiae that harbors many evolutionary conserved proteins. Proteins reach vacuoles via the Vps10-dependent endosomal vacuolar protein sorting pathway, via the alkaline phosphatase (ALP or AP-3) pathway, and via the cytosol-to-vacuole transport (CVT) pathway. A systematic understanding of the cargo spectrum of each pathway is completely lacking. Here, we use quantitative proteomics of purified vacuoles to generate the yeast lysosomal biogenesis map. This dataset harbors information on the cargo-receptor relationship of almost all vacuolar proteins. We map binding motifs of Vps10 and the AP-3 complex and identify a novel cargo of the CVT pathway under nutrient-rich conditions. Our data show how organelle purification and quantitative proteomics can uncover fundamental insights into organelle biogenesis.
溶酶体是细胞中主要的分解细胞器，已被确定为关键的代谢信号中心。许多溶酶体蛋白的突变会产生灾难性的影响，并导致神经退行性疾病、癌症和与年龄相关的疾病。液泡是酿酒酵母中溶酶体的类似物，其中含有许多进化上保守的蛋白质。蛋白质通过 Vps10 依赖性内体液泡蛋白分选途径、碱性磷酸酶 (ALP 或 AP-3) 途径和细胞质到液泡运输 (CVT) 途径到达液泡。每种途径的货物谱的系统理解完全缺乏。在这里，我们使用纯化液泡的定量蛋白质组学生成酵母溶酶体发生图谱。该数据集包含了几乎所有液泡蛋白的货物-受体关系的信息。我们绘制了 Vps10 和 AP-3 复合物的结合基序，并在营养丰富的条件下确定了 CVT 途径的一种新货物。我们的数据表明，细胞器纯化和定量蛋白质组学如何能够揭示细胞器发生的基本见解。
