Thole, Joseph F;
Waudby, Christopher A;
Pielak, Gary J;
(2023)
Disordered proteins mitigate the temperature dependence of site-specific binding free energies.
Journal of Biological Chemistry
, 299
(3)
, Article 102984. 10.1016/j.jbc.2023.102984.
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Abstract
Biophysical characterization of protein-protein interactions involving disordered proteins is challenging. A common simplification is to measure the thermodynamics and kinetics of disordered site binding using peptides containing only the minimum residues necessary. We should not assume, however, that these few residues tell the whole story. Son of sevenless (Sos), a multidomain signaling protein from Drosophila melanogaster, is critical to the mitogen-activated protein kinase pathway, passing an external signal to Ras, which leads to cellular responses. The disordered 55 kDa C-terminal domain of Sos is an auto-inhibitor that blocks guanidine exchange factor activity. Activation requires another protein, Downstream of receptor kinase (Drk), which contains two Src homology 3 (SH3) domains. Here, we utilize nuclear magnetic resonance spectroscopy and isothermal titration calorimetry to quantify the thermodynamics and kinetics of the N-terminal SH3 domain binding to the strongest sites incorporated into the flanking disordered sequences. Comparing these results to those for isolated peptides provides information about how the larger domain affects binding. The affinities of sites on the disordered domain are like those of the peptides at low temperatures but less sensitive to temperature. Our results, combined with observations showing that IDPs become more compact with increasing temperature, suggest a mechanism for this effect.
Type: | Article |
---|---|
Title: | Disordered proteins mitigate the temperature dependence of site-specific binding free energies |
Location: | United States |
Open access status: | An open access version is available from UCL Discovery |
DOI: | 10.1016/j.jbc.2023.102984 |
Publisher version: | https://doi.org/10.1016/j.jbc.2023.102984 |
Language: | English |
Additional information: | This is an open access author accepted manuscript under the CC BY 4.0 license Attribution 4.0 International (https://creativecommons.org/licenses/by/4.0/) |
Keywords: | Drk, Downstream of receptor kinase, IDP, intrinsically disordered protein, IDR, intrinsically disordered region, ITC, isothermal titration calorimetry, NMR, nuclear magnetic resonance, MBD, maltose binding domain, SH3Src homology 3 domains, Sos, Son of sevenless, TEVtobacco etch virus |
UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy |
URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10164632 |
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