論文 - 詳細
| RRC ID | 73961 |
|---|---|
| 著者 | Suzuki T, Kondo C, Kanamori T, Inouye S. |
| タイトル | Video-rate bioluminescence imaging of matrix metalloproteinase-2 secreted from a migrating cell. |
| ジャーナル | PLoS One |
| Abstract |
BACKGROUND:Matrix metalloproteinase-2 (MMP-2) plays an important role in cancer progression and metastasis. MMP-2 is secreted as a pro-enzyme, which is activated by the membrane-bound proteins, and the polarized distribution of secretory and the membrane-associated MMP-2 has been investigated. However, the real-time visualizations of both MMP-2 secretion from the front edge of a migration cell and its distribution on the cell surface have not been reported. METHODOLOGY/PRINCIPAL FINDINGS:The method of video-rate bioluminescence imaging was applied to visualize exocytosis of MMP-2 from a living cell using Gaussia luciferase (GLase) as a reporter. The luminescence signals of GLase were detected by a high speed electron-multiplying charge-coupled device camera (EM-CCD camera) with a time resolution within 500 ms per image. The fusion protein of MMP-2 to GLase was expressed in a HeLa cell and exocytosis of MMP-2 was detected in a few seconds along the leading edge of a migrating HeLa cell. The membrane-associated MMP-2 was observed at the specific sites on the bottom side of the cells, suggesting that the sites of MMP-2 secretion are different from that of MMP-2 binding. CONCLUSIONS:We were the first to successfully demonstrate secretory dynamics of MMP-2 and the specific sites for polarized distribution of MMP-2 on the cell surface. The video-rate bioluminescence imaging using GLase is a useful method to investigate distribution and dynamics of secreted proteins on the whole surface of polarized cells in real time. |
| 巻・号 | 6(9) |
| ページ | e25243 |
| 公開日 | 2011-1-1 |
| DOI | 10.1371/journal.pone.0025243 |
| PII | PONE-D-11-08586 |
| PMID | 21969874 |
| PMC | PMC3182179 |
| MeSH | Cell Membrane / metabolism Cell Movement Disease Progression Gene Expression Regulation, Enzymologic Gene Expression Regulation, Neoplastic HeLa Cells Humans Luminescence Matrix Metalloproteinase 2 / metabolism* Microscopy / methods Microscopy, Fluorescence / methods Neoplasm Metastasis Time Factors Video Recording |
| IF | 2.74 |
| リソース情報 | |
| 遺伝子材料 | pJNC-hMMP2-GLuc (RDB19846) |