ID | 115899 |
著者 |
Kido, Kohki
Proteo-Science Center
Yamanaka, Satoshi
Proteo-Science Center
Nakano, Shogo
University of Shizuoka
Shinohara, Souta
Proteo-Science Center
Nozawa, Akira
Proteo-Science Center
Ito, Sohei
University of Shizuoka
Sawasaki, Tatsuya
Proteo-Science Center
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資料タイプ |
学術雑誌論文
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抄録 | 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.
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掲載誌名 |
eLife
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ISSN | 2050084X
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出版者 | eLife Sciences Publications
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巻 | 9
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開始ページ | e54983
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発行日 | 2020-05-11
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権利情報 | This article is distributed under the terms of the Creative Commons Attribution License(https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use and redistribution provided that the original author and source are credited.
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言語 |
eng
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出版社版
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部局 |
先端酵素学研究所
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