Design of a Highly Selective Quenched Activity-Based Probe and Its Application in Dual Color Imaging Studies of Cathepsin S Activity Localization
- 1 April 2015
- journal article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 137 (14), 4771-4777
- https://doi.org/10.1021/jacs.5b00315
Abstract
The cysteine cathepsins are a group of 11 proteases whose function was originally believed to be the degradation of endocytosed material with a high degree of redundancy. However, it has become clear that these enzymes are also important regulators of both health and disease. Thus, selective tools that can discriminate between members of this highly related class of enzymes will be critical to further delineate the unique biological functions of individual cathepsins. Here we present the design and synthesis of a near-infrared quenched activity-based probe (qABP) that selectively targets cathepsin S which is highly expressed in immune cells. Importantly, this high degree of selectivity is retained both in vitro and in vivo. In combination with a new green-fluorescent pan-reactive cysteine cathepsin qABP we performed dual color labeling studies in bone marrow-derived immune cells and identified vesicles containing exclusively cathepsin S activity. This observation demonstrates the value of our complementary cathepsin probes and provides evidence for the existence of specific localization of cathepsin S activity in dendritic cells.Keywords
Funding Information
- European Research Council (269019)
- National Heart, Lung, and Blood Institute (R01 HL116307)
- National Institute of Biomedical Imaging and Bioengineering (3R01EB005011-06S1, R01 EB005011)
- Technologiestichting STW
- National Institute of General Medical Sciences (R01 GM054051)
- KWF Kankerbestrijding (KUN2009-4402)
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