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High-throughput characterization of protein–protein interactions by reprogramming yeast mating

2017; National Academy of Sciences; Volume: 114; Issue: 46 Linguagem: Inglês

10.1073/pnas.1705867114

1091-6490

David Younger, S. A. Berger, David Baker, Eric Klavins,

Transgenic Plants and Applications

Significance De novo design of protein binders often requires experimental screening to select functional variants from a design library. We have achieved high-throughput, quantitative characterization of protein–protein binding interactions without requiring purified recombinant proteins, by linking interaction strength with yeast mating. Using a next-generation sequencing output, we have characterized protein networks consisting of thousands of pairwise interactions in a single tube and have demonstrated the effect of changing the binding environment. This approach addresses an existing bottleneck in protein binder design by enabling the high-throughput and quantitative characterization of binding strength between designed protein libraries and multiple target proteins in a fully defined environment.