TY - JOUR
T1 - Application of a Sulfoxonium Ylide Electrophile to Generate Cathepsin X-Selective Activity-Based Probes
AU - Mountford, Simon J.
AU - Anderson, Bethany M.
AU - Xu, Bangyan
AU - Tay, Elean S.V.
AU - Szabo, Monika
AU - Hoang, My Linh
AU - Diao, Jiayin
AU - Aurelio, Luigi
AU - Campden, Rhiannon I.
AU - Lindström, Erik
AU - Sloan, Erica K.
AU - Yates, Robin M.
AU - Bunnett, Nigel W.
AU - Thompson, Philip E.
AU - Edgington-Mitchell, Laura E.
N1 - Funding Information:
We thank C. Nowell for maintaining the imaging facilities at the Monash Institute of Pharmaceutical Sciences. MDA-MB-231 cells were a kind gift from Z. Ou, Fudan University, Shanghai Cancer Center. L.E.M. was supported by an Early Career Fellowship from the National Health and Medical Research Council of Australia (NHMRC, GNT1091636), a Grimwade Fellowship funded by the Russell and Mab Grimwade Miegunyah Fund at the University of Melbourne, a DECRA Fellowship from the Australian Research Council (ARC, DE180100418), and by seed grants from Monash University. N.W.B. was supported by grants from the National Institutes of Health (NS102722, DE026806, DK118971) and the United States Department of Defense (W81XWH1810431). HM
Funding Information:
The authors declare the following competing financial interest(s): N.W.B. is a founding scientist of Endosome Therapeutics Inc. Research in N.W.B.'s laboratory is supported in part by Takeda Pharmaceuticals, Inc. Acknowledgments
Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/3/20
Y1 - 2020/3/20
N2 - Cathepsin X/Z/P is cysteine cathepsin with unique carboxypeptidase activity. Its expression is associated with cancer and neurodegenerative diseases, although its roles during normal physiology are still poorly understood. Advances in our understanding of its function have been hindered by a lack of available tools that can specifically measure the proteolytic activity of cathepsin X. We present a series of activity-based probes that incorporate a sulfoxonium ylide warhead, which exhibit improved specificity for cathepsin X compared to previously reported probes. We apply these probes to detect cathepsin X activity in cell and tissue lysates, in live cells and in vivo, and to localize active cathepsin X in mouse tissues by microscopy. Finally, we utilize an improved method to generate chloromethylketones, necessary intermediates for synthesis of acyloxymethylketones probes, by way of sulfoxonium ylide intermediates. In conclusion, the probes presented in this study will be valuable for investigating cathepsin X pathophysiology.
AB - Cathepsin X/Z/P is cysteine cathepsin with unique carboxypeptidase activity. Its expression is associated with cancer and neurodegenerative diseases, although its roles during normal physiology are still poorly understood. Advances in our understanding of its function have been hindered by a lack of available tools that can specifically measure the proteolytic activity of cathepsin X. We present a series of activity-based probes that incorporate a sulfoxonium ylide warhead, which exhibit improved specificity for cathepsin X compared to previously reported probes. We apply these probes to detect cathepsin X activity in cell and tissue lysates, in live cells and in vivo, and to localize active cathepsin X in mouse tissues by microscopy. Finally, we utilize an improved method to generate chloromethylketones, necessary intermediates for synthesis of acyloxymethylketones probes, by way of sulfoxonium ylide intermediates. In conclusion, the probes presented in this study will be valuable for investigating cathepsin X pathophysiology.
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U2 - 10.1021/acschembio.9b00961
DO - 10.1021/acschembio.9b00961
M3 - Article
C2 - 32022538
AN - SCOPUS:85082144574
VL - 15
SP - 718
EP - 727
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 3
ER -