論文 - 詳細
| RRC ID | 72999 |
|---|---|
| 著者 | Kimura Y, Kashima D, Kawahara M. |
| タイトル | A growth-based platform for detecting domain-peptide interactions in the cytoplasm of mammalian cells. |
| ジャーナル | Sci Rep |
| Abstract |
Development of a method for detecting protein-protein interactions (PPIs) in living cells is important for therapeutic drug screening against various diseases including infectious diseases. We have recently developed a method named SOS localization-based interaction screening (SOLIS), in which we designed membrane-anchored and SOS-fused chimeric proteins, whose PPI-dependent association triggers membrane localization of the SOS-fused chimeric protein, activates the Ras/MAPK pathway, and induces cell growth. While SOLIS was able to detect relatively strong PPIs, further sensitivity was required for detecting intracellular endogenous PPIs typically having a micromolar order of dissociation constant (Kd). Here we develop high-sensitive SOLIS (H-SOLIS) that could universally detect PPIs with lower affinities. In order to improve the sensitivity, H-SOLIS introduces a heterodimeric helper interaction, in which addition of a small-molecule helper ligand could accommodate association of the two chimeric proteins and regulate the sensitivity. Four types of domain-peptide interactions having known Kd values are employed to examine the versatility and detection limit of H-SOLIS. Consequently, the heterodimer-inducible helper ligand dramatically enhances detection sensitivity, lowering the detection limit to a ten-micromolar order of Kd. Thus, H-SOLIS could be a platform to detect disease-related domain-peptide interactions for drug discovery screening. |
| 巻・号 | 12(1) |
| ページ | 18028 |
| 公開日 | 2022-10-27 |
| DOI | 10.1038/s41598-022-22770-4 |
| PII | 10.1038/s41598-022-22770-4 |
| PMID | 36302843 |
| PMC | PMC9607845 |
| MeSH | Animals Cytoplasm Ligands Mammals Peptides* / chemistry Protein Interaction Mapping* / methods Recombinant Fusion Proteins Son of Sevenless Proteins |
| IF | 3.998 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | Ba/F3(RCB0805) |