Dataset for analyzing lysosomal proteome using MiSE method

Herein, we develop a novel lysosome-localizable reactive diazirine molecule MDA and demonstrate its enhanced labeling capability in lysosomal microenvironment. Further, we develop a novel microenvironment-specific enrichment (MiSE) strategy for profiling lysosomal proteome, containing MDA-based labeling and affinity enrichment. We demonstrate the effectiveness of MiSE in profiling the lysosomal proteome of living SH-SY5Y cells, identifying 1651 lysosomal proteome, including 126 lysosome-annotated proteins. Furthermore, we utilize MiSE coupled with data-independent acquisition (DIA) analysis to investigate the dynamic changes in the lysosomal proteome in SH-SY5Y cells upon ubiquitin-proteasome system (UPS) inhibition using four proteasome inhibitors. Our results reveal 117 UPS-inhibition related lysosomal proteins, highlighting their involvement in stress response and cell cycle regulation. Notably, we observe distinct proteomic signatures for each inhibitor, suggesting the unique mechanisms of lysosomal response to UPS inhibition.