Imaging specific cell-surface proteolytic activity in single living cells

被引：52

作者：

Hobson, JP

Liu, SH

Rono, B

Leppla, SH

Bugge, TH

机构：

[1] NIAID, Microbial Pathogenesis Sect, NIH, Bethesda, MD 20892 USA

[2] Natl Inst Dent & Craniofacial Res, Proteases & Tissue Remodeling Unit, NIH, Bethesda, MD 20892 USA

[3] Rigshosp, Finsen Lab, DK-2100 Copenhagen O, Denmark

来源：

NATURE METHODS | 2006年 / 3卷 / 04期

关键词：

D O I：

10.1038/NMETH862

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

收藏

页码：259 / 261

页数：3

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