Fig. 5: A2Mcap-based selection of SCHEMA-shuffled Spl proteases.
From: Evolution of protease activation and specificity via alpha-2-macroglobulin-mediated covalent capture
a Schematic of selection strategy for SCHEMA-shuffled proteases using three different substrates and nanopore-based sequencing (b) Sequence analysis of input and output populations. Within each graph, sequences of shuffled variants are depicted as lines coloured by block ID at each position. The number of mappable INC-seq reads is shown for each condition/population. c Flow cytometry of input and output populations probed with the three different A2M substrates used during selection (representative data of n = 2 independent inductions and flow cytometry measurements for 1st round and input library, n = 1 for 2nd round). RCA rolling circle amplification, PCR polymerase chain reaction, MACS magnetic activated cell sorting, FITC Fluorescein isothiocyanate, Strep-PEStreptavidin-Phycoerythrin. Source data is provided as a Source Data file for b and c.
