Proximity proteomics of T-tubules

To identify candidate genes unbiasedly, we employed biotinylation-based proximity proteomics of T-tubules in vivo. We generated a transgenic line carrying an Amph construct fused with miniTurbo (mnTb). We expressed either Amph-mnTb or mnTb alone in muscles, and larvae were reared in the presence or absence of biotin. As a control, LacZ was expressed instead of the mnTb fused constructs. Biotinylated peptides were extracted purified from trypsin-digested samples using Tamavidin 2-REV beads and analyzed by mass spectrometry.