Lysosomal Amino Acid Efflux Assay

Abstract

As the cellular “recycling” organelle, lysosomes break down proteins into amino acids, which are then transported into cytosol for reuse by various amino acid transporters. The transport rate of an amino acid is presumably regulated by cellular conditions such as organelle pH, membrane potential and metabolic states. Because of their intracellular localization and the relative inaccessibility, lysosomal amino acid transporters have been studied largely via indirect measurements. Using lysosome purification and 14C-labeled amino acids, this protocol provides a method to measure the efficiency of specific amino acid transporters on lysosomes., 作为细胞"再循环"细胞器，溶酶体将蛋白质分解成氨基酸，然后将其转运到细胞质中以供各种氨基酸转运蛋白重复使用。 氨基酸的转运速率可能通过细胞条件如细胞器pH，膜电位和代谢状态来调节。 由于它们的细胞内定位和相对不可接近性，溶酶体氨基酸转运蛋白主要通过间接测量进行研究。 使用溶酶体纯化和14 C标记的氨基酸，该协议提供了一种测量特定氨基酸转运蛋白在溶酶体上的效率的方法。