title: Fluorescent tags influence the enzymatic activity and subcellular localization of procaspase-1.
abstract: Subcellular localization studies and life cell imaging approaches usually benefit from fusion-reporter proteins, such as enhanced green fluorescent protein (EGFP) and mCherry to the proteins of interest. However, such manipulations have several risks, including protein misfolding, altered protein shuttling, or functional impairment when compared to the wild-type proteins. Here, we demonstrate altered subcellular distribution and function of the pro-inflammatory enzyme procaspase-1 as a result of fusion with the reporter protein mCherry. Our observations are of central importance to further investigations of subcellular behavior and possible protein-protein interactions of naturally occurring genetic variants of human procaspase-1 which have recently been linked to autoinflammatory disorders.
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Heymann
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Rabe
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Ruß
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Franz
authors.initials: F
authors.lastname: Kapplusch
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Schulze
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Stein
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Winkler
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Hedrich
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
authors.forename: Angela
authors.initials: A
authors.lastname: Rösen-Wolff
authors.affiliation: Department of Pediatrics, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. Electronic address: sigrun.hofmann@uniklinikum-dresden.de.
authors.forename: Sigrun R
authors.initials: SR
authors.lastname: Hofmann
mesh_terms: D020170:Caspase 1 / Q000378:metabolism*; D057075:Enzyme Assays; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D049408:Luminescent Agents / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D000097573:Red Fluorescent Protein
chemical_list: D020170:Caspase 1; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D049408:Luminescent Agents; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein
keywords: CARD; Caspase-1; Fluorescent fusion protein; IL-1β; Inflammasome; mCherry