Quantitative mapping of protein-peptide affinity landscapes using spectrally encoded beads
| Autor: | Brian Baxter, Tanja Kortemme, Kurt S. Thorn, Jagoree Roy, Huy V. Nguyen, Polly M. Fordyce, Björn Harink, Nikhil P Damle, Martha S. Cyert, Kara Brower |
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| Rok vydání: | 2018 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Cell signaling 010405 organic chemistry Globular protein Phosphatase Energy landscape Peptide Computational biology 01 natural sciences 0104 chemical sciences 03 medical and health sciences chemistry Short linear motif Peptide sequence Function (biology) 030304 developmental biology |
| DOI: | 10.1101/306779 |
| Popis: | Transient, regulated binding of globular protein domains to Short Linear Motifs (SLiMs) in disordered regions of other proteins drives cellular signaling. Mapping the energy landscapes of these interactions is essential for deciphering and therapeutically perturbing signaling networks, but is challenging due to their weak affinities. We present a powerful technology, MRBLE-pep, that simultaneously quantifies protein binding to a library of peptides directly synthesized on beads containing unique spectral codes. Using computational modeling and MRBLE-pep, we systematically probe binding of calcineurin (CN), a conserved protein phosphatase essential for the immune response and target of immunosuppressants, to the PxIxIT SLiM. We establish that flanking residues and post- translational modifications critically contribute to PxIxIT-CN affinity, and discover CN-inhibitory peptides with unprecedented affinity and therapeutic potential. The quantitative measurements provided by this approach will improve computational modeling efforts, elucidate a broad range of weak protein-SLiM interactions, and revolutionize our understanding of signaling networks. |
| Databáze: | OpenAIRE |
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