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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
資料タイプ
学術雑誌論文
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.
掲載誌名
eLife
ISSN
2050084X
出版者
eLife Sciences Publications
9
開始ページ
e54983
発行日
2020-05-11
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.
EDB ID
出版社版DOI
出版社版URL
フルテキストファイル
言語
eng
著者版フラグ
出版社版
部局
先端酵素学研究所