title: Video-rate bioluminescence imaging of protein secretion from a living cell.
abstract: We have established a method of video-rate bioluminescence imaging to visualize exocytotic protein secretion from a single living cell using Gaussia luciferase (GLase) as a reporter protein. The luminescence signals produced by the enzymatic reaction of secreted GLase (luciferase) and coelenterazine (luciferin) are detected with an electron-multiplying charge-coupled device camera. An exocytotic event of protein secretion can be visualized using the protein fused to GLase with a time resolution of 30-500 ms. Signal analyses of the bioluminescence video images reveal a number of exocytotic sites, the frequency of exocytotic events, and the amount of secreted protein on a whole cell. Furthermore, the method can distinguish between secreted and cell surface-bound proteins. Our method is a direct approach to investigate the secretion and localization of proteins on the whole surface of living cells.
authors.affiliation: Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Satoshi
authors.initials: S
authors.lastname: Inouye
mesh_terms: D002470:Cell Survival; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D014741:Video Recording / Q000379:methods*
chemical_list: D020778:Matrix Metalloproteinase 2