Proteomics

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Comparison of three enrichment methods of biotinylated peptides or proteins from AirID-CRBN-expressing MM1.S cells

ABSTRACT: MM1.S cells stably expressing AGIA-AirID-CRBN were treated with DMSO or 10 µM pomalidomide in the presence of 10 µM biotin and 5 µM MG132 for 8 h in three biological replicates. The cells were divided into three tubes and lysed. After tryptic digestion, biotinylated peptides were enriched using Tamavidin 2-REV magnetic beads or anti-biotin antibody beads. Biotinylated proteins were enriched using NanoLink streptavidin magnetic beads followed by digestion with Trypsin. The resultant peptides were analyzed by LC-MS/MS.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Hidetaka Kosako 

PROVIDER: PXD028754 | JPOST Repository | Sat Nov 06 00:00:00 GMT 2021

REPOSITORIES: jPOST

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Dataset's files

Source:
Action DRS
210702_Yamanaka_AirID_CRBN_Pom_Tamavidin_FusionIT.pdResult Other
210707_Yamanaka_AirID_CRBN_Pom_Biotin_Ab_FusionIT.pdResult Other
362_Yamanaka_1_DMSO_Tamavidin_FusionIT.raw Raw
364_Yamanaka_4_Pomalidomide_Tamavidin_FusionIT.raw Raw
366_Yamanaka_2_DMSO_Tamavidin_FusionIT.raw Raw
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Publications

A proximity biotinylation-based approach to identify protein-E3 ligase interactions induced by PROTACs and molecular glues.

Yamanaka Satoshi S   Horiuchi Yuto Y   Matsuoka Saya S   Kido Kohki K   Nishino Kohei K   Maeno Mayaka M   Shibata Norio N   Kosako Hidetaka H   Sawasaki Tatsuya T  

Nature communications 20220110 1

Proteolysis-targeting chimaeras (PROTACs) as well as molecular glues such as immunomodulatory drugs (IMiDs) and indisulam are drugs that induce interactions between substrate proteins and an E3 ubiquitin ligases for targeted protein degradation. Here, we develop a workflow based on proximity-dependent biotinylation by AirID to identify drug-induced neo-substrates of the E3 ligase cereblon (CRBN). Using AirID-CRBN, we detect IMiD-dependent biotinylation of CRBN neo-substrates in vitro and identif  ...[more]

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