title: iGLuc: a luciferase-based inflammasome and protease activity reporter.
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)-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.
authors.affiliation: Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Bartok
authors.affiliation: Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Germany.|Department of Internal Medicine III, University Hospital, University of Bonn, Germany.
authors.forename: Franz
authors.initials: F
authors.lastname: Bauernfeind
authors.affiliation: Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Germany.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Khaminets
authors.affiliation: Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Jakobs
authors.affiliation: Institute for Innate Immunity, University Hospital, University of Bonn, Germany.|Division of Infectious Diseases & Immunology, University of Massachusetts Medical School.
authors.forename: Brian
authors.initials: B
authors.lastname: Monks
authors.affiliation: Division of Infectious Diseases & Immunology, University of Massachusetts Medical School.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Fitzgerald
authors.affiliation: Institute for Innate Immunity, University Hospital, University of Bonn, Germany.|Division of Infectious Diseases & Immunology, University of Massachusetts Medical School.
authors.forename: Eicke
authors.initials: E
authors.lastname: Latz
authors.affiliation: Institute for Clinical Chemistry and Pharmacology, Unit for Clinical Biochemistry, University Hospital, University of Bonn, Germany.
authors.forename: Veit
authors.initials: V
authors.lastname: Hornung
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D020170:Caspase 1 / Q000378:metabolism; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism*; D051379:Mice; D014162:Transfection
chemical_list: D058847:Inflammasomes; D053583:Interleukin-1beta; D008156:Luciferases; D020170:Caspase 1