Publication

Visualization of polarized membrane type 1 matrix metalloproteinase activity in live cells by fluorescence resonance energy transfer imaging

Ouyang, M., Lu, S., Li, X-Y., Xu, J., Seong, J., Giepmans, B. N. G., Shyy, J. Y. -J., Weiss, S. J. & Wang, Y., 20-Jun-2008, In : The Journal of Biological Chemistry. 283, 25, p. 17740-17748 9 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • Mingxing Ouyang
  • Shaoying Lu
  • Xiao-Yan Li
  • Jing Xu
  • Jihye Seong
  • Ben N. G. Giepmans
  • John Y. -J. Shyy
  • Stephen J. Weiss
  • Yingxiao Wang

Membrane type 1 matrix metalloproteinase (MT1-MMP) plays a critical role in cancer cell biology by proteolytically remodeling the extracellular matrix. Utilizing fluorescence resonance energy transfer ( FRET) imaging, we have developed a novel biosensor, with its sensing element anchoring at the extracellular surface of cell membrane, to visualize MT1-MMP activity dynamically in live cells with subcellular resolution. Epidermal growth factor (EGF) induced significant FRET changes in cancer cells expressing MT1-MMP, but not in MT1-MMP-deficient cells. EGF-induced FRET changes in MT1-MMP-deficient cells could be restored after reconstituting with wild-type MT1-MMP, but not MMP-2, MMP-9, or inactive MT1-MMP mutants. Deletion of the transmembrane domain in the biosensor or treatment with tissue inhibitor of metalloproteinase-2, a cell-impermeable MT1-MMP inhibitor, abolished the EGF-induced FRET response, indicating that MT1-MMP acts at the cell surface to generate FRET changes. In response to EGF, active MT1-MMP was directed to the leading edge of migrating cells along micropatterned fibronectin stripes, in tandem with the local accumulation of the EGF receptor, via a process dependent upon an intact cytoskeletal network. Hence, the MT1-MMP biosensor provides a powerful tool for characterizing the molecular processes underlying the spatiotemporal regulation of this critical class of enzymes.

Original languageEnglish
Pages (from-to)17740-17748
Number of pages9
JournalThe Journal of Biological Chemistry
Volume283
Issue number25
Publication statusPublished - 20-Jun-2008

    Keywords

  • GROWTH-FACTOR RECEPTOR, 1-MATRIX METALLOPROTEINASE, EXTRACELLULAR-MATRIX, CERVICAL-CARCINOMA, KINASE ACTIVATION, ENDOTHELIAL-CELLS, LIVING CELLS, EGF RECEPTOR, MT1-MMP, SRC

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