フルテキストファイル
著者
Sakurai, Chiye Division of Molecular Biology, School of Life Sciences, Faculty of Medicine, Tottori University / Department of Cell Science, Institute of Biomedical Science, Fukushima Medical University School of Medicine 研究者総覧 KAKEN
Itakura, Makoto Department of Biochemistry, Kitasato University School of Medicine
Kinoshita, Daiki Division of Molecular Biology, School of Life Sciences, Faculty of Medicine, Tottori University
Arai, Seisuke Department of Cell Science, Institute of Biomedical Science, Fukushima Medical University School of Medicine
Hashimoto, Hitoshi Department of Cell Science, Institute of Biomedical Science, Fukushima Medical University School of Medicine
Wada, Ikuo Department of Cell Science, Institute of Biomedical Science, Fukushima Medical University School of Medicine
Hatsuzawa, Kiyotaka Division of Molecular Biology, School of Life Sciences, Faculty of Medicine, Tottori University / Department of Cell Science, Institute of Biomedical Science, Fukushima Medical University School of Medicine 研究者総覧 KAKEN
抄録
SNAP-23 is a plasma membrane-localized soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNARE) involved in Fc receptor (FcR)-mediated phagocytosis. However, the regulatory mechanism underlying its function remains elusive. Using phosphorylation-specific antibodies, SNAP-23 was found to be phosphorylated at Ser95 in macrophages. To understand the role of this phosphorylation, we established macrophage lines overexpressing the nonphosphorylatable S95A or the phosphomimicking S95D mutation. The efficiency of phagosome formation and maturation was severely reduced in SNAP-23-S95D–overexpressing cells. To examine whether phosphorylation at Ser95 affected SNAP-23 structure, we constructed intramolecular Förster resonance energy transfer (FRET) probes of SNAP-23 designed to evaluate the approximation of the N termini of the two SNARE motifs. Interestingly, a high FRET efficiency was detected on the membrane when the S95D probe was used, indicating that phosphorylation at Ser95 caused a dynamic structural shift to the closed form. Coexpression of IκB kinase (IKK) 2 enhanced the FRET efficiency of the wild-type probe on the phagosome membrane. Furthermore, the enhanced phagosomal FRET signal in interferon-γ–activated macrophages was largely dependent on IKK2, and this kinase mediated a delay in phagosome-lysosome fusion. These results suggested that SNAP-23 phosphorylation at Ser95 played an important role in the regulation of SNARE-dependent membrane fusion during FcR-mediated phagocytosis.
出版者
American Society for Cell Biology
資料タイプ
学術雑誌論文
外部リンク
ISSN
10591524
EISSN
19394586
掲載誌名
MOLECULAR BIOLOGY OF THE CELL
最新掲載誌名
MOLECULAR BIOLOGY OF THE CELL
29
14
開始ページ
1753
終了ページ
1762
発行日
2018-07-15
出版者DOI
著者版フラグ
出版社版
著作権表記
© 2018 Sakurai et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
掲載情報
Sakurai Chiye, Itakura Makoto, Kinoshita Daiki, et al. Phosphorylation of SNAP-23 at Ser95 causes a structural alteration and negatively regulates Fc receptor-mediated phagosome formation and maturation in macrophages. MOLECULAR BIOLOGY OF THE CELL. 2018. 29(14). 1753-1762. doi:10.1091/mbc.E17-08-0523
部局名
医学部・医学系研究科・医学部附属病院
言語
英語
Web of Science Key ut
WOS:000444415400008