MRBLE-pep measurements reveal accurate binding affinities for B56, a PP2A regulatory subunit
| Autor: | Jamin B. Hein, Martha S. Cyert, Polly M. Fordyce |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
Environmental Engineering QD71-142 Chemistry Protein subunit Peptide Isothermal titration calorimetry Computational biology Protein phosphatase 2 Affinities Industrial and Manufacturing Engineering Article Wide dynamic range Short linear motif Analytical chemistry Binding selectivity |
| Zdroj: | ACS Measurement Science Au, Vol 1, Iss 2, Pp 56-64 (2021) ACS Meas Sci Au Hein, J B, Cyert, M S & Fordyce, P M 2021, ' MRBLE-pep Measurements Reveal Accurate Binding Affinities for B56, a PP2A Regulatory Subunit ', ACS Measurement Science Au, vol. 1, no. 2, pp. 56-64 . https://doi.org/10.1021/acsmeasuresciau.1c00008 |
| Popis: | Signal transduction pathways rely on dynamic interactions between protein globular domains and short linear motifs (SLiMs). The weak affinities of these interactions are essential to allow fast rewiring of signaling pathways and downstream responses, but pose technical challenges for interaction detection and measurement. We recently developed a technique (MRBLE-pep) that leverages spectrally encoded hydrogel beads to measure binding affinities between a single protein and 48 different peptide sequences in a single small volume. In prior work, we applied it to map the binding specificity landscape between calcineurin and the PxIxIT SLiM (Nguyen et al. 2019). Here, using peptide sequences known to bind the PP2A regulatory subunit B56, we systematically compare affinities measured by MRBLE-pep or isothermal calorimetry (ITC) and confirm that MRBLE-pep accurately quantifies relative affinity over a wide dynamic range while using a fraction of the material required for traditional methods such as ITC. |
| Databáze: | OpenAIRE |
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