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ChemComm
Page 4 of 4
DOI: 10.1039/C6CC04030F
COMMUNICATION
Journal Name
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new list of candidates was generated after MS analysis. In
these two experiments, a total of 11 candidates were detected
through the probe-modified peptides, which also revealed the
modification site on the target proteins (Figure S9).
Benefiting from BTC-ABPP, finding the modification site of
the probe can reveal its possible mechanisms of action. For
example, CTSC, one candidate identified both in the trypsin
digestion and in the pepsin digestion, was modified by
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Canertinib probe at Cys234. Many CTSC inhibitors are
irreversible inhibitors that covalently bind to Cys234 residue in
the catalytic site19, indicating Canertinib as a potential CTSC
inhibitor. Thus we developed CTSC enzymatic assay according
to the previous report(Figure 5a).20 Canertinib exhibited dose-
dependent inhibition to CTSC activity, with an IC50 of 0.12 µM
(Figure 5b). CTSC is involved in the activation of granule-
associated serine proteases and is also associated with many
inflammatoryand autoimmune diseases such as Rheumatoid
Arthritis and Chronic Obstructive Pulmonary Disease (COPD).18
Most known CTSC inhibitors are analogs of the dipeptide
substrates, but few can enter clinical trials due to poor
metabolic stability. Now Canertinib can be used as a new
parent nucleus to develop novel CTSC inhibitors which will
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In conclusion, we have developed a new method based on
ABPP technology by converting the reactants from proteins to
peptides, and changing the manipulation system from
biochemistry to chemistry. This method can both improve the
efficiency of click conjugation reaction and reduce false-
positive targets. Besides, under the guidance of the
information encoded by the probe-modified peptide (ID of
target and the modification site), BTC-ABPP can effectively
discriminate the functional target from other unrelated
candidates, and accelerate the target validation process. Thus,
the BTC-ABPP method should be a good alternative to the
known ABPP technology, and will benefit the inevitable
discovery of biologically improtant targets and facilitate the
mechanism study.
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We thank Dr. Wanzhong He for assistance with the electron
micrographs. We acknowledge the National High Technology
Project 973 (2011CB504300). We thank Prof. Zhenhua Zhang
(China Agricultural University) for providing several azide
reagents (National Key Technologies R&D Program of China,
2015BAK45B01, CAU).
20. Thong, B.; Pilling, J.; Ainscow, E.; Beri, R.; Unitt, J., J. Biomol.
Screen. 2011, 16, 36.
Notes and references
4 | J. Name., 2012, 00, 1-3
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