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  1. C100 医学部/医学系研究科
  2. C100b 紀要
  3. Nagoya journal of medical science
  4. 83(3)

Screening of novel Midkine binding protein by BioID2-based proximity labeling

https://doi.org/10.18999/nagjms.83.3.495
2124988f-a05d-4a1c-8ba9-1e59f82313b5
名前 / ファイル ライセンス アクション
08_Komata.pdf 08_Komata.pdf (9.5 MB)
Item type itemtype_ver1(1)
公開日 2021-09-16
タイトル
タイトル Screening of novel Midkine binding protein by BioID2-based proximity labeling
言語 en
著者 Komata, Yosuke

× Komata, Yosuke

en Komata, Yosuke

Search repository
Tsubota, Shoma

× Tsubota, Shoma

en Tsubota, Shoma

Search repository
Sakamoto, Kazuma

× Sakamoto, Kazuma

en Sakamoto, Kazuma

Search repository
Ikematsu, Shinya

× Ikematsu, Shinya

en Ikematsu, Shinya

Search repository
Kadomatsu, Kenji

× Kadomatsu, Kenji

en Kadomatsu, Kenji

Search repository
アクセス権
アクセス権 open access
アクセス権URI http://purl.org/coar/access_right/c_abf2
キーワード
主題Scheme Other
主題 MK
キーワード
主題Scheme Other
主題 BioID2
キーワード
主題Scheme Other
主題 IGFBP2
内容記述
内容記述 Midkine (MK), a heparin-binding growth factor, is associated with the poor prognosis of the pediatric tumor, neuroblastoma. MK would be a druggable target as many studies showed inhibition of its function in various cancers suppressed tumor developments. To establish the therapy targeting MK, identification of its binding partners, and elucidation of its intracellular signaling are needed. It was reported that exogenous MK induced phosphorylation of ribosomal protein S6 (RPS6) downstream of mTOR signaling. Using RPS6 phosphorylation as a marker of MK response, we searched for MK reactive cell lines. We found that MK cell lines expressing less MK tended to respond better to MK. Next, using an MK reactive neuroblastoma cell line, MK-knocked down SH-SY5Y cells, we employed a proximity-dependent biotin identification method, which was invented to evaluate protein-protein interactions by biotinylation. We confirmed that secreted MK fused to the biotin ligase BioID2 (MK-BioID2) was able to biotinylate proteins from the cells. Biotinylated proteins were identified by liquid chromatography-mass spectrometry analyses. Twenty five proteins were found to be overlapped after three independent experiments, among which insulin-like growth binding protein 2 (IGFBP2) was further analyzed. IGFBP2 was indeed detected with immunoblotting after streptavidin pull down of MK-BioID2 labeled cell extract of MK-knocked down SH-SY5Y cells. Our study suggests that the BioID2 method is useful to identify binding partners of growth factors.
言語 en
内容記述タイプ Abstract
内容記述
内容記述 This is an Open Access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. To view the details of this license, please visit (http://creativecommons.org/licenses/by-nc-nd/4.0/).
言語 en
内容記述タイプ Other
出版者
言語 en
出版者 Nagoya University Graduate School of Medicine, School of Medicine
言語
言語 eng
資源タイプ
資源タイプresource http://purl.org/coar/resource_type/c_6501
タイプ departmental bulletin paper
出版タイプ
出版タイプ VoR
出版タイプResource http://purl.org/coar/version/c_970fb48d4fbd8a85
ID登録
ID登録 10.18999/nagjms.83.3.495
ID登録タイプ JaLC
関連情報
関連タイプ isVersionOf
関連識別子
識別子タイプ URI
関連識別子 https://www.med.nagoya-u.ac.jp/medlib/nagoya_j_med_sci/833.html
収録物識別子
収録物識別子タイプ PISSN
収録物識別子 0027-7622
収録物識別子
収録物識別子タイプ EISSN
収録物識別子 2186-3326
書誌情報 en : Nagoya Journal of Medical Science

巻 83, 号 3, p. 495-508, 発行日 2021-08
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