Fig. 6: Characterisation of Spl protease chimeras.
From: Evolution of protease activation and specificity via alpha-2-macroglobulin-mediated covalent capture
a Flow cytometry of parent Spl proteases and chimeras with ID of variable block 5 position indicated in red (representative data of n = 3 independent inductions and flow cytometry measurements). b SDS-PAGE of E. coli-purified parent and chimera Spl proteins used for in vitro characterization (native N-termini generated via SUMO-tag removal strategy as described in Methods section Protein expression and purification, result of n = 1 protein purification). c Activity measurements (pseudo-first order kinetics) of purified Spl parent proteases and chimeras measured at 0.5–2 μM enzyme and 2.5–5 μM fluorogenic peptide substrates (Ac: acetyl-; AMC: 7-Amino-4-methylcoumarin; n = 3 independent measurements of the same protein purification, mean with SEM). Source data are provided as a Source Data file for panels a, b and c. FITC fluorescein isothiocyanate, Strep-PE streptavidin-phycoerythrin, MW molecular weight, kDa kilodalton.
