論文 - 詳細
| RRC ID | 62848 |
|---|---|
| 著者 | Kido K, Yamanaka S, Nakano S, Motani K, Shinohara S, Nozawa A, Kosako H, Ito S, Sawasaki T. |
| タイトル | AirID, a novel proximity biotinylation enzyme, for analysis of protein-protein interactions. |
| ジャーナル | Elife |
| Abstract |
Proximity biotinylation based on Escherichia coli BirA enzymes such as BioID (BirA*) and TurboID is a key technology for identifying proteins that interact with a target protein in a cell or organism. However, there have been some improvements in the enzymes that are used for that purpose. Here, we demonstrate a novel BirA enzyme, AirID (ancestral BirA for proximity-dependent biotin identification), which was designed de novo using an ancestral enzyme reconstruction algorithm and metagenome data. AirID-fusion proteins such as AirID-p53 or AirID-IκBα indicated biotinylation of MDM2 or RelA, respectively, in vitro and in cells, respectively. AirID-CRBN showed the pomalidomide-dependent biotinylation of IKZF1 and SALL4 in vitro. AirID-CRBN biotinylated the endogenous CUL4 and RBX1 in the CRL4CRBN complex based on the streptavidin pull-down assay. LC-MS/MS analysis of cells that were stably expressing AirID-IκBα showed top-level biotinylation of RelA proteins. These results indicate that AirID is a novel enzyme for analyzing protein-protein interactions. |
| 巻・号 | 9 |
| 公開日 | 2020-5-11 |
| DOI | 10.7554/eLife.54983 |
| PII | 54983 |
| PMID | 32391793 |
| PMC | PMC7302878 |
| MeSH | Biotin / chemistry Biotin / metabolism Biotinylation Carbon-Nitrogen Ligases / chemistry* Carbon-Nitrogen Ligases / genetics Cell Survival Escherichia coli / enzymology Escherichia coli / genetics Escherichia coli Proteins / chemistry* Escherichia coli Proteins / genetics HEK293 Cells Humans Mutation Protein Engineering* Protein Interaction Mapping Protein Interaction Maps Recombinant Fusion Proteins / chemistry* Recombinant Fusion Proteins / genetics Repressor Proteins / chemistry* Repressor Proteins / genetics |
| IF | 7.08 |
| 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 | 29 |
| 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 | 293T(RCB2202) |