The goal of this project is to identify changes of the lysosomal surface proteome upon the induction of lysosomal vacuolation. 293T cells stably expressing LAMP1-GFP-TurboID or Lyso-TurboID were treated with a small molecule apilimod or DMSO to trigger lysosomal vacuolation, after which the lysosomal surface proteins were labeled with biotin by TurboID. The biotinylated proteins by Lyso-TurboID were purified by streptavidin beads for mass spectrometry.