Reference - Detail
| RRC ID | 86381 |
|---|---|
| Author | Yamada K, Yamanaka S, Yamakoshi H, Kohyama A, Iwabuchi Y, Kosako H, Sawasaki T. |
| Title | In-cell proximity target validation methods for heterobifunctional molecules with CRBN- or VHL-binder using AirID. |
| Journal | Commun Biol |
| Abstract |
Heterobifunctional molecules, such as proteolysis-targeting and autophagy-targeting chimera, represent new drug concepts. They are composed of two protein binders that can induce proximity interactions between two proteins and protein catalysis. Currently, cereblon (CRBN)- and von Hippel-Lindau (VHL)-binders with thalidomide- and VH032-backbones are widely used as E3 ligase binders. Here, we developed a method to validate proteins that interact with heterobifunctional molecules in cells using AirID, a proximity biotinylation enzyme. Interactome of target proteins was validated for six heterobifunctional molecules. ThBD-AirID, a fusion of the thalidomide-binding domain (ThBD) of CRBN and AirID, effectively biotinylated the target proteins. AirID fused to full-length VHL also exhibited highly effective biotinylation. Heterobifunctional molecules with the same target binder but different E3 binders showed different proximity interactome profiles in cells. Analysis using ThBD-AirID revealed a nuclear interaction between androgen receptor and ARV-110. AirID-fused ThBD and VHL could be useful for validating the heterobifunctional molecular interactome in cells. |
| Volume | 8(1) |
| Pages | 1323 |
| Published | 2025-8-30 |
| DOI | 10.1038/s42003-025-08761-x |
| PII | 10.1038/s42003-025-08761-x |
| PMID | 40885760 |
| PMC | PMC12398604 |
| MeSH | Adaptor Proteins, Signal Transducing* / genetics Adaptor Proteins, Signal Transducing* / metabolism Biotinylation HEK293 Cells Humans Protein Binding Thalidomide / metabolism Ubiquitin-Protein Ligases* / metabolism Von Hippel-Lindau Tumor Suppressor Protein* / genetics Von Hippel-Lindau Tumor Suppressor Protein* / metabolism |
| IF | 4.165 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 9 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | B16F10(RCB2630) LNCap.FGC(RCB2144) |
| DNA material | pCAGGS (RDB08938) |