Genetically encoded fluorescent biosensors for live-cell imaging of MT1-MMP protease activity.
The proteolytic activity of Membrane-type 1 Matrix Metalloproteinase (MT1-MMP) is crucial for cancer cell invasion and metastasis. To visualize the protease activity of MT1-MMP with high spatiotemporal resolution at the extracellular plasma membrane surface of live cancer cells, a genetically encoded fluorescent biosensor of MT1-MMP has been developed. Here we describe the design principles of the MT1-MMP biosensor, the characterization of the MT1-MMP biosensor in vitro, and the live-cell imaging protocol used to visualize MT1-MMP activity in mammalian cells. We also provide brief guidelines for observing MT1-MMP subcellular activity by fluorescence resonance energy transfer (FRET) in a cell migration assay.
Department of Bioengineering, University of Illinois at Urbana-Champaign, Champaign, IL, USA.
Mingxing
M
Ouyang
Shaoying
S
Lu
Yingxiao
Y
Wang
D000595:Amino Acid Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D031541:Fluorescence Resonance Energy Transfer; D005818:Genetic Engineering; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism
D010455:Peptides; D053511:Matrix Metalloproteinase 14