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Literature DB >> 16554829

Imaging specific cell-surface proteolytic activity in single living cells.

John P Hobson¹, Shihui Liu, Birgitte Rønø, Stephen H Leppla, Thomas H Bugge.

Abstract

We describe a simple, sensitive and noninvasive assay that uses nontoxic, reengineered anthrax toxin-beta-lactamase fusion proteins with altered protease cleavage specificity to visualize specific cell-surface proteolytic activity in single living cells. The assay could be used to specifically image endogenous cell-surface furin, urokinase plasminogen activator and metalloprotease activity. We have adapted the assay for fluorescence microscopy, flow cytometry and fluorescent plate reader formats, and it is amenable for automation and high-throughput analysis.

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Year: 2006 PMID： 16554829 DOI： 10.1038/nmeth862

Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547

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Cited

26 in total

1. Dissecting the urokinase activation pathway using urokinase-activated anthrax toxin.

Authors: Shihui Liu; Thomas H Bugge; Arthur E Frankel; Stephen H Leppla
Journal: Methods Mol Biol Date: 2009

Review 2. Measuring enzyme activity in single cells.

Authors: Michelle L Kovarik; Nancy L Allbritton
Journal: Trends Biotechnol Date: 2011-02-11 Impact factor: 19.536

Review 3. Designing inhibitors of anthrax toxin.

Authors: Ekaterina M Nestorovich; Sergey M Bezrukov
Journal: Expert Opin Drug Discov Date: 2014-01-22 Impact factor: 6.098

4. Inhibition of anthrax protective antigen outside and inside the cell.

Authors: Marina V Backer; Vimal Patel; Brian T Jehning; Kevin P Claffey; Vladimir A Karginov; Joseph M Backer
Journal: Antimicrob Agents Chemother Date: 2006-10-30 Impact factor: 5.191

5. Inhibition of tumor angiogenesis by the matrix metalloproteinase-activated anthrax lethal toxin in an orthotopic model of anaplastic thyroid carcinoma.

Authors: Randall W Alfano; Stephen H Leppla; Shihui Liu; Thomas H Bugge; Janelle M Ortiz; Terry C Lairmore; Nicholas S Duesbery; Ian C Mitchell; Fiemu Nwariaku; Arthur E Frankel
Journal: Mol Cancer Ther Date: 2010-01-06 Impact factor: 6.261

Imaging specific cell-surface proteolytic activity in single living cells.

1. Dissecting the urokinase activation pathway using urokinase-activated anthrax toxin.

Review 2. Measuring enzyme activity in single cells.

Review 3. Designing inhibitors of anthrax toxin.

4. Inhibition of anthrax protective antigen outside and inside the cell.

5. Inhibition of tumor angiogenesis by the matrix metalloproteinase-activated anthrax lethal toxin in an orthotopic model of anaplastic thyroid carcinoma.

6. Probing enzymatic activity inside living cells using a nanowire-cell "sandwich" assay.

7. Characterization of a Chinese hamster ovary cell mutant having a mutation in elongation factor-2.

8. Imaging proprotein convertase activities and their regulation in the implanting mouse blastocyst.

9. Anthrax toxin uptake by primary immune cells as determined with a lethal factor-beta-lactamase fusion protein.

10. Quantitative high-throughput screening identifies inhibitors of anthrax-induced cell death.