Proximity-dependent TurboID labeling screen on the LAMTOR complex

ABSTRACT: This proximity-dependent labeling screen was set up to investigate the “proxisome” (i.e. the sum of proteins in the labeling diameter around a bait protein) of the LAMTOR complex. Furthermore, we aimed at understanding the recruitment dynamics of known and unknown proteins to and from the LAMTOR complex upon changing environmental conditions. Therefore, cells were exposed to three different conditions: Full medium (FM), amino acid and growth factor starvation (EBSS) and replenishment of amino acid and growth factors after starvation (HF). To achieve that, we generated stable cell lines expressing the biotin ligase TurboID fused to two LAMTOR subunits (LAMTOR3 and LAMTOR5) as well as to a lysosomal (TMEM192) and cytosolic (nuclear exclusion sequence NES) control. After induction of the biotin-ligase reaction in living cells, biotinylated proteins were extracted from cell lysates via streptavidin beads and further processed for mass spectrometry analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Markus Hartl  

LAB HEAD: Prof. Manuela Baccarini

PROVIDER: PXD080046 | Pride | 2026-07-10

REPOSITORIES: Pride