Chapter title |
Genetically Encoded Fluorescent Biosensors for Live-Cell Imaging of MT1-MMP Protease Activity.
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Chapter number | 13 |
Book title |
Fluorescent Protein-Based Biosensors
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Published in |
Methods in molecular biology, January 2014
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DOI | 10.1007/978-1-62703-622-1_13 |
Pubmed ID | |
Book ISBNs |
978-1-62703-621-4, 978-1-62703-622-1
|
Authors |
Mingxing Ouyang, Shaoying Lu, Yingxiao Wang, Ouyang M, Lu S, Wang Y, Ouyang, Mingxing, Lu, Shaoying, Wang, Yingxiao |
Abstract |
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. |
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