徳島大学 教育・研究者情報データベース(EDB)

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著作: Kido Kohki/Yamanaka Satoshi/Nakano Shogo/[茂谷 康]/Shinohara Souta/Nozawa Akira/[小迫 英尊]/Ito Sohei/Sawasaki Tatsuya/AirID, a Novel Proximity Biotinylation Enzyme, for Analysis of Protein-Protein Interactions/[eLife]

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EID
366537
EOID
983661
Map
0
LastModified
2020年7月31日(金) 14:26:56
Operator
大家 隆弘
Avail
TRUE
Censor
0
Owner
小迫 英尊
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種別 必須 学術論文(審査論文)
言語 必須 英語
招待 推奨
審査 推奨
カテゴリ 推奨
共著種別 推奨
学究種別 推奨
組織 推奨
著者 必須
  1. (英) Kido Kohki
    役割 任意
    貢献度 任意
    学籍番号 推奨
  2. (英) Yamanaka Satoshi
    役割 任意
    貢献度 任意
    学籍番号 推奨
  3. (英) Nakano Shogo
    役割 任意
    貢献度 任意
    学籍番号 推奨
  4. 茂谷 康([徳島大学.先端酵素学研究所.基幹研究部門])
    役割 任意
    貢献度 任意
    学籍番号 推奨
  5. (英) Shinohara Souta
    役割 任意
    貢献度 任意
    学籍番号 推奨
  6. (英) Nozawa Akira
    役割 任意
    貢献度 任意
    学籍番号 推奨
  7. 小迫 英尊([徳島大学.先端酵素学研究所.基幹研究部門])
    役割 任意
    貢献度 任意
    学籍番号 推奨
  8. (英) Ito Sohei
    役割 任意
    貢献度 任意
    学籍番号 推奨
  9. (英) Sawasaki Tatsuya
    役割 任意
    貢献度 任意
    学籍番号 推奨
題名 必須

(英) AirID, a Novel Proximity Biotinylation Enzyme, for Analysis of Protein-Protein Interactions
副題 任意
要約 任意

(英) Proximity biotinylation based on 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 CRL4 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.
キーワード 推奨
発行所 推奨
誌名 必須 eLife(eLife Sciences Publications, Ltd.)
(eISSN: 2050-084X)
ISSN 任意 2050-084X
ISSN: 2050-084X (eISSN: 2050-084X)
Title: eLife
Title(ISO): Elife
Publisher: eLife Sciences Publications
 (NLM Catalog)   (Scopus)   (CrossRef)  (Scopus information is found. [need login])
巻 必須 9
号 必須 e54983
頁 必須 ---
都市 任意
年月日 必須 2020年 5月 11日
URL 任意
DOI 任意 10.7554/eLife.54983    (→Scopusで検索)
PMID 任意 32391793    (→Scopusで検索)
CRID 任意
WOS 任意
Scopus 任意
評価値 任意
被引用数 任意
指導教員 推奨
備考 任意

  1. (英) Article.ELocationID: 10.7554/eLife.54983

  2. (英) Article.ELocationID: e54983

  3. (英) Article.PublicationTypeList.PublicationType: Journal Article

  4. (英) OtherAbstract.AbstractText: Proteins in a cell need to interact with each other to perform the many tasks required for organisms to thrive. A technique called proximity biotinylation helps scientists to pinpoint the identity of the proteins that partner together. It relies on attaching an enzyme (either BioID or TurboID) to a protein of interest; when a partner protein comes in close contact with this construct, the enzyme can attach a chemical tag called biotin to it. The tagged proteins can then be identified, revealing which molecules interact with the protein of interest. Although BioID and TurboID are useful tools, they have some limitations. Experiments using BioID take more than 16 hours to complete and require high levels of biotin to be added to the cells. TurboID is more active than BioID and is able to label proteins within ten minutes. However, under certain conditions, it is also more likely to be toxic for the cell, or to make mistakes and tag proteins that do not interact with the protein of interest. To address these issues, Kido et al. developed AirID, a new enzyme for proximity biotinylation. Experiments were then conducted to test how well AirID would perform, using proteins of interest whose partners were already known. These confirm that AirID was able to label partner proteins in human cells; compared with TurboID, it was also less likely to mistakenly tag non-partners or to kill the cells, even over long periods. The results by Kido et al. demonstrate that AirID is suitable for proximity biotinylation experiments in cells. Unlike BioID and TurboID, the enzyme may also have the potential to be used for long-lasting experiments in living organisms, since it is less toxic for cells over time.

  5. (英) KeywordList.Keyword: A. thaliana

  6. (英) KeywordList.Keyword: BioID

  7. (英) KeywordList.Keyword: ancestral sequence reconstruction

  8. (英) KeywordList.Keyword: biochemistry

  9. (英) KeywordList.Keyword: cell biology

  10. (英) KeywordList.Keyword: cell-free

  11. (英) KeywordList.Keyword: chemical biology

  12. (英) KeywordList.Keyword: enzymatic engineering

  13. (英) KeywordList.Keyword: human

  14. (英) KeywordList.Keyword: protein-protein interaction

  15. (英) KeywordList.Keyword: proximity labeling

  16. (英) CoiStatement: KK, SY, SN, KM, SS, AN, HK, SI, TS No competing interests declared