title: A novel approach for reliable detection of cathepsin S activities in mouse antigen presenting cells.
abstract: Cathepsin S (CTSS) is a eukaryotic protease mostly expressed in professional antigen presenting cells (APCs). Since CTSS activity regulation plays a role in the pathogenesis of various autoimmune diseases like multiple sclerosis, atherosclerosis, Sjögren's syndrome and psoriasis as well as in cancer progression, there is an ongoing interest in the reliable detection of cathepsin S activity. Various applications have been invented for specific detection of this enzyme. However, most of them have only been shown to be suitable for human samples, do not deliver quantitative results or the experimental procedure requires technical equipment that is not commonly available in a standard laboratory. We have tested a fluorogen substrate, Mca-GRWPPMGLPWE-Lys(Dnp)-DArg-NH2, that has been described to specifically detect CTSS activities in human APCs for its potential use for mouse samples. We have modified the protocol and thereby offer a cheap, easy, reproducible and quick activity assay to detect CTSS activities in mouse APCs. Since most of basic research on CTSS is performed in mice, this method closes a gap and offers a possibility for reliable and quantitative CTSS activity detection that can be performed in almost every laboratory.
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Alex
authors.initials: A
authors.lastname: Steimle
authors.affiliation: Interfacultary Institute of Biochemistry, University of Tübingen, Germany.
authors.forename: Hubert
authors.initials: H
authors.lastname: Kalbacher
authors.affiliation: Institute of Radiology, University of Tübingen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Maurer
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Beifuss
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Annika
authors.initials: A
authors.lastname: Bender
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Schäfer
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Ricarda
authors.initials: R
authors.lastname: Müller
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany.
authors.forename: Ingo B
authors.initials: IB
authors.lastname: Autenrieth
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Tübingen, Germany. Electronic address: julia-stefanie.frick@med.uni-tuebingen.de.
authors.forename: Julia-Stefanie
authors.initials: JS
authors.lastname: Frick
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology / Q000382:microbiology; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004927:Escherichia coli Infections / Q000201:enzymology* / Q000276:immunology / Q000382:microbiology; D005456:Fluorescent Dyes / Q000378:metabolism*; D006868:Hydrolysis; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D018345:Mice, Knockout; D010455:Peptides / Q000378:metabolism*; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D015853:Cysteine Proteinase Inhibitors; D005456:Fluorescent Dyes; D010455:Peptides; D002403:Cathepsins; C028217:cathepsin S; D007930:Leucine; C024974:E 64
keywords: Autoimmune disease; Cathepsin S; Enzyme activity; IBD; Method; Mouse; Multiple sclerosis