iGLuc: a luciferase-based inflammasome and protease activity reporter.
Nat Methods
; 10(2): 147-154, 2013 Feb.
Article
en En
| MEDLINE
| ID: mdl-23291722
ABSTRACT
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)-1ß-Gaussia luciferase (iGLuc) fusion construct, in which pro-IL-1ß-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.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Técnicas Biosensibles
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Interleucina-1beta
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Inflamasomas
/
Luciferasas
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Nat Methods
Asunto de la revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Año:
2013
Tipo del documento:
Article
País de afiliación:
Alemania