Pepstatin-Based Probes for Photoaffinity Labeling of Aspartic Proteases and Application to Target Identification

Aspartic proteases are a small class of proteases implicated in a wide variety of human diseases. Covalent chemical probes for photoaffinity labeling (PAL) of these proteases are underdeveloped. We here report a full on-resin synthesis of clickable PAL probes based on the natural product inhibitor p...

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Bibliographic Details
Published in:ACS chemical biology 2023-04, Vol.18 (4), p.686-692
Main Authors: Chen, Suyuan, Liang, Chunguang, Li, Hongli, Yu, Weimeng, Prothiwa, Michaela, Kopczynski, Dominik, Loroch, Stefan, Fransen, Marc, Verhelst, Steven H. L.
Format: Article
Language:English
Subjects:
Aspartic Acid Endopeptidases - chemistry
Cathepsin D - chemistry
Diazomethane
Humans
Pepstatins - chemistry
Pepstatins - pharmacology
Photoaffinity Labels - chemistry
Sequestosome-1 Protein - chemistry
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Summary:Aspartic proteases are a small class of proteases implicated in a wide variety of human diseases. Covalent chemical probes for photoaffinity labeling (PAL) of these proteases are underdeveloped. We here report a full on-resin synthesis of clickable PAL probes based on the natural product inhibitor pepstatin incorporating a minimal diazirine reactive group. The position of this group in the inhibitor determines the labeling efficiency. The most effective probes sensitively detect cathepsin D, a biomarker for breast cancer, in cell lysates. Moreover, through chemical proteomics experiments and deep learning algorithms, we identified sequestosome-1, an important player in autophagy, as a direct interaction partner and substrate of cathepsin D.
ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.2c00946