|
Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Life Science / Faculty of Advanced Life Science >
Peer-reviewed Journal Articles, etc >
Monitoring the caspase cascade in single apoptotic cells using a three-color fluorescent protein substrate
| Title: | Monitoring the caspase cascade in single apoptotic cells using a three-color fluorescent protein substrate |
| Authors: | Sun, Fan Browse this author | | Mikuni, Shintaro Browse this author | | Kinjo, Masataka Browse this author |
| Keywords: | FCCS | | Caspase inhibition | | Multi color | | Morphological change |
| Issue Date: | 14-Jan-2011 |
| Publisher: | Elsevier |
| Journal Title: | Biochemical and Biophysical Research Communications |
| Volume: | 404 |
| Issue: | 2 |
| Start Page: | 706 |
| End Page: | 710 |
| Publisher DOI: | 10.1016/j.bbrc.2010.12.047 |
| PMID: | 21156159 |
| Abstract: | Fluorescence cross-correlation spectroscopy (FCCS) reveals information about the spatiotemporal coincidence of two spectrally well-defined fluorescent molecules in a small observation area at the level of single-molecule sensitivity. To simultaneously evaluate the activities of caspase-3 and caspase-9, we constructed a chimeral protein that consisted of tandemly fused enhanced cyan fluorescent protein (ECFP), monomeric red fluorescent protein (mCherry) and monomeric yellow fluorescent protein (Venus). In HeLa cell lysates, a combination of tumor necrosis factor-α (TNF-α)- and cycloheximide (CHX-)-induced apoptosis was monitored. In this, decreases of cross-correlation amplitudes were observed between ECFP and mCherry and between mCherry and Venus. Moreover, time-dependent monitoring of single cells revealed decreases in the cross-correlation amplitudes between ECFP and mCherry and between mCherry and Venus before morphologic changes were observed by laser scanning fluorescence microscopy (LSM). Thus, our method could predict the fate of the cell in the early apoptotic stage. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/44926 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
|
Submitter: 金城 政孝
|
