Quantitative mapping of protein-peptide affinity landscapes using spectrally encoded beads
| Autor: | Kurt S. Thorn, Polly M. Fordyce, Naomi R. Latorraca, Scott A. Longwell, Björn Harink, Brian Baxter, Jagoree Roy, Tanja Kortemme, Nikhil P Damle, Martha S. Cyert, Kara Brower, Huy V. Nguyen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cell signaling QH301-705.5 Globular protein Science Structural Biology and Molecular Biophysics Chemical biology microfluidics Computational biology Plasma protein binding phophatase specificity Binding Competitive General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Biochemistry and Chemical Biology Peptide Library Phosphoprotein Phosphatases Humans Short linear motif Amino Acid Sequence Protein Interaction Maps Biology (General) chemistry.chemical_classification 030102 biochemistry & molecular biology General Immunology and Microbiology General Neuroscience Calcineurin Molecular biophysics protein specificity Proteins Hydrogels General Medicine Models Theoretical Affinities Microspheres protein-peptide interactions 030104 developmental biology chemistry Structural biology spectral encoding Medicine Peptides Protein Processing Post-Translational Algorithms Research Article Human Protein Binding |
| Zdroj: | eLife eLife, Vol 8 (2019) |
| ISSN: | 2050-084X |
| 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 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 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 discover that flanking residues and post-translational modifications critically contribute to PxIxIT-CN affinity and identify CN-binding peptides based on multiple scaffolds with a wide range of affinities. The quantitative biophysical data 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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