| RRC ID |
86381
|
| 著者 |
Yamada K, Yamanaka S, Yamakoshi H, Kohyama A, Iwabuchi Y, Kosako H, Sawasaki T.
|
| タイトル |
In-cell proximity target validation methods for heterobifunctional molecules with CRBN- or VHL-binder using AirID.
|
| ジャーナル |
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.
|
| 巻・号 |
8(1)
|
| ページ |
1323
|
| 公開日 |
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
|
| リソース情報 |
| ヒト・動物細胞 |
B16F10(RCB2630)
LNCap.FGC(RCB2144) |
| 遺伝子材料 |
pCAGGS (RDB08938) |
