TY - JOUR
T1 - Application of an Activity-Based Probe to Determine Proteolytic Activity of Cell Surface Cathepsin G by Mass Cytometry Data Acquisition
AU - Gärtner, Fabian
AU - Knippschild, Uwe
AU - Burster, Timo
N1 - Funding Information:
FG is a member of the International Graduate School of Molecular Medicine (IGradU) at Ulm University, which is funded by the DFG under the acronym “GSC270”. We gratefully thank Marcin Sienczyk (Faculty of Chemistry, Wroclaw University of Technology, Wroclaw, Poland) for providing MARS-116-Bt, structure, and Suc-VPF. Furthermore, we thank Sarah Warth from the Core Facility of Cytometry (Ulm University, Ulm, Germany) for measuring different samples by CyTOF. T.B. was funded by the Nazarbayev University Faculty-Development Competitive Research Grants Program, reference: 280720FD1907.
Publisher Copyright:
Copyright © 2020 American Chemical Society.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020
Y1 - 2020
N2 - During an immune response, cathepsin G (CatG) takes on the role of adaptive and innate immunity and the outcome depends on the localization of CatG. Soluble, cell surface-bound, or intracellular CatG is also responsible for pathophysiology conditions. We applied the activity-based probe MARS116-Bt to mass cytometry by time-of-flight to analyze CatG activity on the cell surface of immune cells. The phosphonate warhead of MARS116-Bt binds covalently to the serine amino acid residue S195 of the catalytic center and thereby CatG activity can be detected. This method contributes to observing the activation or inhibition status of cells during pathogenesis of diseases and enables accurate data acquisition from complex biological samples with a vast panel of cell subset markers in a single-cell resolution.
AB - During an immune response, cathepsin G (CatG) takes on the role of adaptive and innate immunity and the outcome depends on the localization of CatG. Soluble, cell surface-bound, or intracellular CatG is also responsible for pathophysiology conditions. We applied the activity-based probe MARS116-Bt to mass cytometry by time-of-flight to analyze CatG activity on the cell surface of immune cells. The phosphonate warhead of MARS116-Bt binds covalently to the serine amino acid residue S195 of the catalytic center and thereby CatG activity can be detected. This method contributes to observing the activation or inhibition status of cells during pathogenesis of diseases and enables accurate data acquisition from complex biological samples with a vast panel of cell subset markers in a single-cell resolution.
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U2 - 10.1021/acsomega.0c04092
DO - 10.1021/acsomega.0c04092
M3 - Article
AN - SCOPUS:85096099591
VL - 5
SP - 28233
EP - 28238
JO - ACS Omega
JF - ACS Omega
SN - 2470-1343
IS - 43
ER -