Proteomic analysis of silenced cathepsin B expression suggests non-proteolytic cathepsin B functionality

Cathepsin B (CTSB) is a lysosomal endo- and exopeptidase that is also secreted in high amounts by malignant and non-malignant cells. We determined the effect of CTSB on the tumor cell secretome by shRNA-mediated silencing of CTSB mRNA expression and subsequent proteomic LC-MS/MS analysis of the cell...

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Hauptverfasser: Sigloch, Florian Christoph (VerfasserIn) , Knopf, Julia Daniela (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 12 August 2016
In: Biochimica et biophysica acta. Molecular cell research
Year: 2016, Jahrgang: 1863, Heft: 11, Pages: 2700-2709
ISSN:1879-2596
DOI:10.1016/j.bbamcr.2016.08.005
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.bbamcr.2016.08.005
Verlag, lizenzpflichtig, Volltext: http://www.sciencedirect.com/science/article/pii/S0167488916302105
Volltext
Verfasserangaben:Florian Christoph Sigloch, Julia Daniela Knopf, Juliane Weißer, Alejandro Gomez-Auli, Martin Lothar Biniossek, Agnese Petrera, Oliver Schilling
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Zusammenfassung:Cathepsin B (CTSB) is a lysosomal endo- and exopeptidase that is also secreted in high amounts by malignant and non-malignant cells. We determined the effect of CTSB on the tumor cell secretome by shRNA-mediated silencing of CTSB mRNA expression and subsequent proteomic LC-MS/MS analysis of the cell supernatants. We identified significant protein changes of 17 secreted or shed proteins. Notably, we found a general reduction in protein abundance of ADAM10 substrates and lysosomal proteins. We corroborated reduced amounts of soluble ADAM10 (sADAM10) and soluble APP (sAPP) in the two cancer cell lines MDA-MB-231 and U2OS by immunoblotting. Interestingly, reductions in sADAM10 and sAPP could be reversed by re-introducing a catalytically inactive variant of CTSB, suggesting a formerly unknown non-catalytic function of the protease.
Beschreibung:Gesehen am 07.05.2020
Beschreibung:Online Resource
ISSN:1879-2596
DOI:10.1016/j.bbamcr.2016.08.005