著作: [茂谷 康]/[小迫 英尊]/BioID screening of biotinylation sites using the avidin-like protein Tamavidin 2-REV identifies global interactors of stimulator of interferon genes (STING)./[The Journal of Biological Chemistry]
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種別 | 必須 | 学術論文(審査論文) | |||
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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 | ||||
題名 | 必須 |
(英) BioID screening of biotinylation sites using the avidin-like protein Tamavidin 2-REV identifies global interactors of stimulator of interferon genes (STING). |
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副題 | 任意 | ||||
要約 | 任意 |
(英) Stimulator of interferon genes (STING) mediates cytosolic DNA-induced innate immune signaling via membrane trafficking. Global identification of proteins that spatiotemporally interact with STING will provide a better understanding of its trafficking mechanisms and of STING signaling pathways. Proximity-dependent biotin identification (BioID) is a powerful technology to identify physiologically relevant protein-protein interactions in living cells. However, biotinylated peptides are rarely detected in the conventional BioID method, which uses streptavidin beads to pull-down biotinylated proteins, because the biotin-streptavidin interaction is too strong. As a result, only non-biotinylated peptides are identified, which cannot be distinguished from peptides of non-specifically pull-downed proteins. Here, we developed a simple method to efficiently and specifically enrich biotinylated peptides using Tamavidin 2-REV, an engineered avidin-like protein with reversible biotin-binding capability. Using RAW264.7 macrophages stably expressing TurboID-fused STING, we identified and quantified >4,000 biotinylated peptides of STING-proximal proteins. Various endoplasmic reticulum-associated proteins were biotinylated in unstimulated cells, and STING activation caused biotinylation of many proteins located in the Golgi and endosomes. These proteins included those known to interact with activated STING, such as TANK-binding kinase 1 (TBK1), several palmitoyl transferases, and p62/sequestosome 1 (SQSTM1). Furthermore, interferon-induced transmembrane protein 3 (IFITM3), an endolysosome-localized antiviral protein, bound to STING at the late activation stage. These dynamic interaction profiles will provide detailed insights into STING signaling; we propose that our approach using Tamavidin 2-REV would be useful for BioID-based and other biotinylation-based peptide identification methods. |
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キーワード | 推奨 | ||||
発行所 | 推奨 | ||||
誌名 | 必須 |
The Journal of Biological Chemistry([The American Society for Biochemistry and Molecular Biology])
(pISSN: 0021-9258, eISSN: 1083-351X)
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巻 | 必須 | 295 | |||
号 | 必須 | 32 | |||
頁 | 必須 | 11174 11183 | |||
都市 | 任意 | ||||
年月日 | 必須 | 2020年 8月 7日 | |||
URL | 任意 | ||||
DOI | 任意 | 10.1074/jbc.RA120.014323 (→Scopusで検索) | |||
PMID | 任意 | 32554809 (→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 |
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