Consequences of inducing intrinsic disorder in a high-affinity protein-protein interaction.

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Papadakos, Grigorios and Sharma, Amit and Lancaster, Lorna E. and Bowen, Rebecca and Kaminska, Renata and Leech, Andrew P. and Walker, Daniel and Redfield, Christina and Kleanthous, Colin (2015) Consequences of inducing intrinsic disorder in a high-affinity protein-protein interaction. Journal of the American Chemical Society, 137 (16). 5252–5255. ISSN 1520-5126 (https://doi.org/10.1021/ja512607r)

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Abstract

The kinetic and thermodynamic consequences of intrinsic disorder in protein–protein recognition are controversial. We address this by inducing one partner of the high-affinity colicin E3 rRNase domain–Im3 complex (Kd ≈ 10–12 M) to become an intrinsically disordered protein (IDP). Through a variety of biophysical measurements, we show that a single alanine mutation at Tyr507 within the hydrophobic core of the isolated colicin E3 rRNase domain causes the enzyme to become an IDP (E3 rRNaseIDP). E3 rRNaseIDP binds stoichiometrically to Im3 and forms a structure that is essentially identical to the wild-type complex. However, binding of E3 rRNaseIDP to Im3 is 4 orders of magnitude weaker than that of the folded rRNase, with thermodynamic parameters reflecting the disorder-to-order transition on forming the complex. Critically, pre-steady-state kinetic analysis of the E3 rRNaseIDP–Im3 complex demonstrates that the decrease in affinity is mostly accounted for by a drop in the electrostatically steered association rate. Our study shows that, notwithstanding the advantages intrinsic disorder brings to biological systems, this can come at severe kinetic and thermodynamic cost.

ORCID iDs

Papadakos, Grigorios, Sharma, Amit, Lancaster, Lorna E., Bowen, Rebecca, Kaminska, Renata, Leech, Andrew P., Walker, Daniel ORCID logoORCID: https://orcid.org/0000-0002-4206-2942, Redfield, Christina and Kleanthous, Colin;
  • Item type:Article
    ID code:86564
    Dates:
    Date
    Event
    29 April 2015
    Published
    17 April 2015
    Published Online
    17 April 2015
    Accepted
    11 December 2014
    Submitted
    Notes:Copyright © 2022 American Chemical Society. Consequences of Inducing Intrinsic Disorder in a High-Affinity Protein–Protein Interaction Grigorios Papadakos, Amit Sharma, Lorna E. Lancaster, Rebecca Bowen, Renata Kaminska, Andrew P. Leech, Daniel Walker, Christina Redfield, and Colin Kleanthous Journal of the American Chemical Society 2015 137 (16), 5252-5255 DOI: 10.1021/ja512607r
    Subjects:Science > Chemistry
    Science > Microbiology
    Department:Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
    Depositing user: Pure Administrator
    Date deposited:24 Aug 2023 10:43
    Last modified:11 Nov 2024 14:01
    URI:https://strathprints.strath.ac.uk/id/eprint/86564
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