Substrate specificity studies of the cysteine peptidases falcipain-2 and falcipain-3 from Plasmodium falciparum and demonstration of their kininogenase activity

► Analysis of FP-2 and FP-3 specificity requirements using FRET peptides as substrates. ► Despite high sequence identity and structural similarity between FP-2 and FP-3, noteworthy differences in substrate specificity were observed. ► Recombinant falcipain-2 and falcipain-3 hydrolyzed a fluorescent kinin related peptide. ► Falcipains excised Met-Lys-BK, Lys-BK and BK from the fluorogenic HK-related sequence.