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Methods Mol Biol. 2014;1071:163-74. doi: 10.1007/978-1-62703-622-1_13.

Genetically encoded fluorescent biosensors for live-cell imaging of MT1-MMP protease activity.

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  • 1Department of Bioengineering, University of Illinois at Urbana-Champaign, Champaign, IL, USA.

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.

[PubMed - indexed for MEDLINE]
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