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Selective imaging of cathepsin L in breast cancer by fluorescent activity-based probes

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Abstract

Cysteine cathepsins normally function in the lysosomal degradation system where they are critical for the maintenance of cellular homeostasis and the MHC II immune response, and have been found to have major roles in several diseases and in tumor progression. Selective visualization of individual protease activity within a complex proteome is of major importance to establish their roles in both normal and tumor cells, thereby facilitating our understanding of the regulation of proteolytic networks. A generally accepted means to monitor protease activity is the use of small molecule substrates and activity-based probes. However, there are eleven human cysteine cathepsins, with a few of them displaying overlapping substrate specificity, making the development of small molecules that selectively target a single cathepsin very challenging. Here, we utilized HyCoSuL, a positional scanning substrate approach, to develop a highly-selective fluorogenic substrate and activity-based probe for monitoring cathepsin L activity in the breast cancer cell line MDA-MB-231. Use of this probe enabled us to distinguish the activity of cathepsin L from that of other cathepsins, particularly cathepsin B, which is abundant and ubiquitously expressed in normal and transformed cell types. We found that cathepsin L localization in MDA-MB-231 cells greatly overlaps with that of cathepsin B, however, several cathepsin L-rich lysosomes lacked cathepsin B activity. Overall, these studies demonstrate that HyCoSuL-derived small molecule probes are valuable tools to image cathepsin L activity in living cells. This approach thus enables evaluation of cathepsin L function in tumorigenesis and is applicable to other cysteine cathepsins.

Graphical abstract: Selective imaging of cathepsin L in breast cancer by fluorescent activity-based probes

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Publication details

The article was received on 03 Oct 2017, accepted on 15 Jan 2018 and first published on 16 Jan 2018


Article type: Edge Article
DOI: 10.1039/C7SC04303A
Citation: Chem. Sci., 2018, Advance Article
  • Open access: Creative Commons BY license
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    Selective imaging of cathepsin L in breast cancer by fluorescent activity-based probes

    M. Poreba, W. Rut, M. Vizovisek, K. Groborz, P. Kasperkiewicz, D. Finlay, K. Vuori, D. Turk, B. Turk, G. S. Salvesen and M. Drag, Chem. Sci., 2018, Advance Article , DOI: 10.1039/C7SC04303A

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