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Limited proteolysis was performed by enzymatic digestion at 37 C adding bovine trypsin (Sigma-Aldrich, Italy) or proteinase K (Sigma-Aldrich, Italy) to protein extract at a ratio 1:100 enzyme/substrate (w/w)

To overcome this limitation, we adapted our cell-based autoprocessing assay for high-throughput screening (HTS) using an amplified luminescent proximity homogeneous assay ELISA (AlphaLISA) to quantify autoprocessing efficiency 41