| 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. | |
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MedLine Citation:
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PMID: 21519891 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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-domain FLIM. Image analysis was carried out both with polar plots and phase differential enhancement. Phase differential enhancement, which is similar to phase suppression, is shown to facilitate the differentiation between different conformations of the MT1-MMP biosensor in live cells when the lifetime differences are small. FLIM carried out in differential enhancement or phase suppression modes, requires only two acquired phase images, and permits rapid imaging of the activity of MT1-MMP in live cells. |
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Authors:
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John Paul Eichorst; He Huang; Robert M Clegg; Yingxiao Wang |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. Date: 2011-04-26 |
Journal Detail:
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Title: Journal of fluorescence Volume: 21 ISSN: 1573-4994 ISO Abbreviation: J Fluoresc Publication Date: 2011 Jul |
Date Detail:
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Created Date: 2011-07-27 Completed Date: 2011-12-29 Revised Date: 2013-04-29 |
Medline Journal Info:
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Nlm Unique ID: 9201341 Medline TA: J Fluoresc Country: Netherlands |
Other Details:
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Languages: eng Pagination: 1763-77 Citation Subset: IM |
Copyright Information:
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© Springer Science+Business Media, LLC 2011 |
Affiliation:
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Center of Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Biosensing Techniques* Cell Line, Tumor Fluorescence Resonance Energy Transfer HeLa Cells Humans Matrix Metalloproteinase 14 / chemistry, metabolism* Microscopy, Fluorescence |
| Grant Support | |
ID/Acronym/Agency:
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CA139272/CA/NCI NIH HHS; HL098472/HL/NHLBI NIH HHS; K02 HL109142/HL/NHLBI NIH HHS; NS063405/NS/NINDS NIH HHS; R01 HL098472/HL/NHLBI NIH HHS; R01 HL098472-01/HL/NHLBI NIH HHS; R01 HL098472-02/HL/NHLBI NIH HHS; R01 HL098472-03/HL/NHLBI NIH HHS; R21 CA139272-01/CA/NCI NIH HHS; R21 CA139272-02/CA/NCI NIH HHS; R21 CA139272-02S1/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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EC 3.4.24.80/Matrix Metalloproteinase 14 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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