Phase differential enhancement of FLIM to distinguish FRET components of a biosensor for monitoring molecular activity of Membrane Type 1 Matrix Metalloproteinase in live cells.

ABSTRACT: Fluorescence lifetime-resolved imaging microscopy (FLIM) has been used to monitor the enzymatic activity of a proteolytic enzyme, Membrane Type 1 Matrix Metalloproteinase (MT1-MMP), with a recently developed FRET-based biosensor in vitro and in live HeLa and HT1080 cells. MT1-MMP is a collagenaise that is involved in the destruction of extra-cellular matrix (ECM) proteins, as well as in various cellular functions including migration. The increased expression of MT1-MMP has been positively correlated with the invasive potential of tumor cells. However, the precise spatiotemporal activation patterns of MT1-MMP in live cells are still not well-established. The activity of MT1-MMP was examined with our biosensor in live cells. Imaging of live cells was performed with full-field frequency-domai

SUBMITTER: Eichorst JP 

PROVIDER: S-EPMC3637990 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature