Fig. 1 | Nature Communications

Fig. 1

From: Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries

Fig. 1

Yeast-surface display of APPI-3M. a Schematic drawing of the pairwise selective screen using the YSD system. A naive library of mutated APPI-3M variants was displayed on the yeast cell surface and presented to pairs of proteases. Each protease in the pair (denoted A or B) was labeled with a different fluorescent dye—Alexa Fluor-650 or Alexa Fluor-488 (represented by green and blue stars, respectively). b Pairwise selective screen. Flow cytometry sorting was used to screen the library to isolate APPI-3M variants, with enhanced selectivity toward each of the four serine proteases (Meso: mesotrypsin; KLK6; Anionic: anionic trypsin; Cationic: cationic trypsin). In each sort, two variant populations were collected inside the black gates, yielding sorted library populations of protease-selective APPI-3M variants. Green and blue colors represent a high and low cell densities, respectively

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