2. Academic Validation
  3. Activity-Based Imaging of Acid Ceramidase in Living Cells

Activity-Based Imaging of Acid Ceramidase in Living Cells

  • J Am Chem Soc. 2019 May 15;141(19):7736-7742. doi: 10.1021/jacs.8b11687.
Yadira F Ordóñez 1,  José Luís Abad 1,  Mazen Aseeri 1,  Josefina Casas 1 2,  Virginie Garcia 3,  Mireia Casasampere 1,  Edward H Schuchman 4,  Thierry Levade 3 5,  Antonio Delgado 1 6,  Gemma Triola 7,  Gemma Fabrias 1 2
Affiliations

Affiliations

  • 1 Research Unit on Bioactive Molecules (RUBAM), Department of Biological Chemistry , Institute for Advanced Chemistry of Catalonia (IQAC-CSIC) , Jordi Girona, 18 , 08034 Barcelona , Spain.
  • 2 Liver and Digestive Diseases Networking Biomedical Research Centre (CIBER-EHD) , 28029 Madrid , Spain.
  • 3 INSERM, UMR1037 CRCT , 31037 Toulouse , France.
  • 4 Department of Genetics and Genomic Sciences , Icahn School of Medicine at Mount Sinai , New York 10029 , United States.
  • 5 Laboratoire de Biochimie Métabolique , Institut Fédératif de Biologie, CHU Purpan , 31300 Toulouse , France.
  • 6 Unit of Pharmaceutical Chemistry (Associated Unit to CSIC), Department of Pharmacology, Toxicology and Medicinal Chemistry, Faculty of Pharmacy and Food Sciences , University of Barcelona , 08028 Barcelona , Spain.
  • 7 Chemical Biology group, Department of Biological Chemistry , Institute for Advanced Chemistry of Catalonia (IQAC-CSIC) , Jordi Girona, 18 , 08034 Barcelona , Spain.
PMID: 31030513 DOI: 10.1021/jacs.8b11687
Abstract

Acid Ceramidase (AC) hydrolyzes ceramides into sphingoid bases and fatty acids. The Enzyme is overexpressed in several types of Cancer and Alzheimer's disease, and its genetic defect causes different incurable disorders. The availability of a method for the specific visualization of catalytically active AC in intracellular compartments is crucial for diagnosis and follow-up of therapeutic strategies in diseases linked to altered AC activity. This work was undertaken to develop activity-based probes for the detection of AC. Several analogues of the AC inhibitor SABRAC were synthesized and found to act as very potent (two-digit nM range) irreversible AC inhibitors by reaction with the active site Cys143. Detection of active AC in cell-free systems was achieved either by using fluorescent SABRAC analogues or by Click Chemistry with an azide-substituted analogue. The compound affording the best features allowed the unprecedented labeling of active AC in living cells.

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