Document Detail

iGLuc: a luciferase-based inflammasome and protease activity reporter.
MedLine Citation:
PMID:  23291722     Owner:  NLM     Status:  MEDLINE    
Measurement of protease activity in living cells or organisms remains a challenging task. We here present a transgene-encoded biosensor that reports the proteolytic activity of caspase-1 in the course of inflammasome activation and that of other proteases in a highly sensitive and specific manner. This protease reporter is based on the biological activity of a pro-interleukin (IL)-1β-Gaussia luciferase (iGLuc) fusion construct, in which pro-IL-1β-dependent formation of protein aggregates renders GLuc enzyme inactive. Cleavage leads to monomerization of this biosensor protein, resulting in a strong gain in luciferase activity. Exchange of the canonical caspase-1 cleavage site in this reporter construct allows the generation of protease biosensors with additional specificities. The high sensitivity, signal-to-background ratio and specificity of the iGLuc system renders it a useful tool to study proteolytic events in mouse and human cells at high throughput and to monitor protease activity in mice in vivo.
Eva Bartok; Franz Bauernfeind; Maria G Khaminets; Christopher Jakobs; Brian Monks; Katherine A Fitzgerald; Eicke Latz; Veit Hornung
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2013-01-06
Journal Detail:
Title:  Nature methods     Volume:  10     ISSN:  1548-7105     ISO Abbreviation:  Nat. Methods     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-01-30     Completed Date:  2013-03-28     Revised Date:  2014-08-14    
Medline Journal Info:
Nlm Unique ID:  101215604     Medline TA:  Nat Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  147-54     Citation Subset:  IM    
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MeSH Terms
Biosensing Techniques / methods*
Caspase 1 / metabolism
Cell Line
Enzyme Activation
Inflammasomes / metabolism*
Interleukin-1beta / metabolism*
Luciferases / metabolism*
Grant Support
243046//European Research Council
Reg. No./Substance:
0/Inflammasomes; 0/Interleukin-1beta; EC 1.13.12.-/Luciferases; EC 1

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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