論文 - 詳細
| RRC ID | 68665 |
|---|---|
| 著者 | Sakurai C, Hashimoto H, Nakanishi H, Arai S, Wada Y, Sun-Wada GH, Wada I, Hatsuzawa K. |
| タイトル | SNAP-23 regulates phagosome formation and maturation in macrophages. |
| ジャーナル | Mol Biol Cell |
| Abstract |
Synaptosomal associated protein of 23 kDa (SNAP-23), a plasma membrane-localized soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE), has been implicated in phagocytosis by macrophages. For elucidation of its precise role in this process, a macrophage line overexpressing monomeric Venus-tagged SNAP-23 was established. These cells showed enhanced Fc receptor-mediated phagocytosis. Detailed analyses of each process of phagocytosis revealed a marked increase in the production of reactive oxygen species within phagosomes. Also, enhanced accumulation of a lysotropic dye, as well as augmented quenching of a pH-sensitive fluorophore were observed. Analyses of isolated phagosomes indicated the critical role of SNAP-23 in the functional recruitment of the NADPH oxidase complex and vacuolar-type H(+)-ATPase to phagosomes. The data from the overexpression experiments were confirmed by SNAP-23 knockdown, which demonstrated a significant delay in phagosome maturation and a reduction in uptake activity. Finally, for analyzing whether phagosomal SNAP-23 entails a structural change in the protein, an intramolecular Förster resonance energy transfer (FRET) probe was constructed, in which the distance within a TagGFP2-TagRFP was altered upon close approximation of the N-termini of its two SNARE motifs. FRET efficiency on phagosomes was markedly enhanced only when VAMP7, a lysosomal SNARE, was coexpressed. Taken together, our results strongly suggest the involvement of SNAP-23 in both phagosome formation and maturation in macrophages, presumably by mediating SNARE-based membrane traffic. |
| 巻・号 | 23(24) |
| ページ | 4849-63 |
| 公開日 | 2012-12-1 |
| DOI | 10.1091/mbc.E12-01-0069 |
| PII | mbc.E12-01-0069 |
| PMID | 23087210 |
| PMC | PMC3521691 |
| MeSH | Animals Blotting, Western Cell Line Fluorescence Resonance Energy Transfer Humans Luminescent Proteins / genetics Luminescent Proteins / metabolism Lysosomes / metabolism Macrophages / cytology Macrophages / metabolism* Mice Microscopy, Confocal NADPH Oxidases / metabolism Phagocytosis Phagosomes / metabolism* Protein Binding Qa-SNARE Proteins / metabolism Qb-SNARE Proteins / genetics Qb-SNARE Proteins / metabolism* Qc-SNARE Proteins / genetics Qc-SNARE Proteins / metabolism* R-SNARE Proteins / metabolism RNA Interference Reactive Oxygen Species / metabolism Vacuolar Proton-Translocating ATPases / metabolism |
| IF | 3.791 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | F1000Research |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | J774.1 |