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A novel approach for reliable detection of cathepsin S activities in mouse antigen presenting cells.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2016 May; Vol. 432, pp. 87-94. Date of Electronic Publication: 2016 Feb 18. - Publication Year :
- 2016
-
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.<br /> (Copyright © 2016. Published by Elsevier B.V.)
- Subjects :
- Animals
Antigen-Presenting Cells drug effects
Antigen-Presenting Cells immunology
Antigen-Presenting Cells microbiology
Cathepsins antagonists & inhibitors
Cathepsins deficiency
Cathepsins genetics
Cells, Cultured
Cysteine Proteinase Inhibitors pharmacology
Escherichia coli Infections immunology
Escherichia coli Infections microbiology
Hydrolysis
Leucine analogs & derivatives
Leucine pharmacology
Mice, Knockout
Reproducibility of Results
Substrate Specificity
Time Factors
Antigen-Presenting Cells enzymology
Cathepsins metabolism
Escherichia coli Infections enzymology
Fluorescent Dyes metabolism
Peptides metabolism
Spectrometry, Fluorescence
Subjects
Details
- Language :
- English
- ISSN :
- 1872-7905
- Volume :
- 432
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 26899824
- Full Text :
- https://doi.org/10.1016/j.jim.2016.02.015