Video-rate bioluminescence imaging of matrix metalloproteinase-2 secreted from a migrating cell

• Published in: PloS one Vol. 6; no. 9; p. e25243
• Main Authors: Suzuki, Takahiro, Kondo, Chihiro, Kanamori, Takao, Inouye, Satoshi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.09.2011; Public Library of Science (PLoS)
• Subjects: Biochemistry; Biology; Bioluminescence; Cameras; Cancer; Cancer metastasis; Cell adhesion & migration; Cell Membrane - metabolism; Cell Movement; Cell surface; Charge coupled devices; Dentistry; Disease Progression; Exocytosis; Fusion protein; Gaussia; Gelatinase A; Gene expression; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; HeLa Cells; Humans; Imaging; Luciferase; Luminescence; Matrix metalloproteinase; Matrix Metalloproteinase 2 - metabolism; Medical research; Medicine; Metalloproteinase; Metastases; Microscopy - methods; Microscopy, Fluorescence - methods; Neoplasm Metastasis; Protein binding; Proteins; Real time; Rodents; Secretion; Time Factors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0025243

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. 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. 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.