RRC ID 81308
Author Yamanaka S, Horiuchi Y, Matsuoka S, Kido K, Nishino K, Maeno M, Shibata N, Kosako H, Sawasaki T.
Title A proximity biotinylation-based approach to identify protein-E3 ligase interactions induced by PROTACs and molecular glues.
Journal Nat Commun
Abstract 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 identify biotinylated peptides of well-known neo-substrates by mass spectrometry with high specificity and selectivity. Additional analyses reveal ZMYM2 and ZMYM2-FGFR1 fusion protein-responsible for the 8p11 syndrome involved in acute myeloid leukaemia-as CRBN neo-substrates. Furthermore, AirID-DCAF15 and AirID-CRBN biotinylate neo-substrates targeted by indisulam and PROTACs, respectively, suggesting that this approach has the potential to serve as a general strategy for characterizing drug-inducible protein-protein interactions in cells.
Volume 13(1)
Pages 183
Published 2022-1-10
DOI 10.1038/s41467-021-27818-z
PII 10.1038/s41467-021-27818-z
PMID 35013300
PMC PMC8748630
MeSH Adaptor Proteins, Signal Transducing / genetics* Adaptor Proteins, Signal Transducing / metabolism Biological Assay* Biotinylation Cell Line, Tumor DNA-Binding Proteins / genetics* DNA-Binding Proteins / metabolism Gene Expression Regulation HEK293 Cells Hepatocytes / cytology Hepatocytes / drug effects Hepatocytes / metabolism* Humans Immunologic Factors / pharmacology Intracellular Signaling Peptides and Proteins / genetics Intracellular Signaling Peptides and Proteins / metabolism Lymphocytes / cytology Lymphocytes / drug effects Lymphocytes / metabolism* Neurons / cytology Neurons / drug effects Neurons / metabolism Protein Binding Protein Interaction Mapping Proteolysis / drug effects Receptor, Fibroblast Growth Factor, Type 1 / genetics Receptor, Fibroblast Growth Factor, Type 1 / metabolism Substrate Specificity Sulfonamides / pharmacology Transcription Factors / genetics* Transcription Factors / metabolism Ubiquitin-Protein Ligases / genetics* Ubiquitin-Protein Ligases / metabolism
Resource
DNA material pCAGGS (RDB08938) CSII-CMV-MCS-IRES2-Bsd (RDB04385) pCMV-VSV-G-RSV-Rev (RDB04393) pCAG-HIVgp (RDB04394)