A novel approach for reliable detection of cathepsin S activities in mouse antigen presenting cells

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• 作者：Alex Steimle^a ; Hubert Kalbacher^b ; Andreas Maurer^c ; Brigitte Beifuss^a ; Annika Bender^a ; Andrea Schä ; fer^a ; Ricarda Mü ; ller^a ; Ingo B. Autenrieth^a ; Julia-Stefanie Frick^a ; ^{julia-stefanie.frick@med.uni-tuebingen.de" class="auth_mail" title="E-mail the corresponding author}
• 关键词：APCs ; antigen presenting cells ; BMDCs ; bone marrow derived dendritic cells ; CTSB ; cathepsin B ; CTSD ; cathepsin D ; CTSL ; cathepsin L ; CTSS ; cathepsin S ; rh ; recombinant human ; PMGLP ; Mca-GRWPPMGLPWE-Lys(Dnp)-DArg-NH2 ; rm ; recombinant mouse ; RT ; room temperature
• 刊名：Journal of Immunological Methods
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：432
• 期：Complete
• 页码：87-94
• 全文大小：750 K

文摘

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.