Extended Data Fig. 4: Assessment of covalent bond formation between NNPI-C10 and CTSS.
From: Antibody–peptide conjugates deliver covalent inhibitors blocking oncogenic cathepsins
a) Fluorescence and coomassie blue staining of gel loaded with CTSS alone or incubated with AF488-derivatized NNPI-C10. Different concentrations of E64 were tested to assess if NNPI-C10 could be displaced from CTSS. Images of 1 out of 2 independent experiments represented. b) Molecular structure of NNPI-C10-NC, differing from C10 because of a single bond replacing the double bond in the Michael acceptor. c) Intact mass spectrometry analysis of CTSS in complex with NNPI-C10 (red peaks) compared to CTSS alone (black peaks), performed in denaturing conditions.
