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Engineering Order and Cooperativity in a Disordered Protein

Munshi, Sneha and Subramanian, Sandhyaa and Ramesh, Samyuktha and Golla, Hemashree and Kalivarathan, Divakar and Kulkarni, Madhurima and Campos, Luis A and Sekhar, Ashok and Naganathan, Athi N (2019) Engineering Order and Cooperativity in a Disordered Protein. In: BIOCHEMISTRY, 58 (19). pp. 2389-2397.

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Official URL: https://doi.org/10.1021/acs.biochem.9b00182

Abstract

Structural disorder in proteins arises from a complex interplay between weak hydrophobicity and unfavorable electrostatic interactions. The extent to which the hydrophobic effect contributes to the unique and compact native state of proteins is, however, confounded by large compensation between multiple entropic and energetic terms. Here we show that protein structural order and cooperativity arise as emergent properties upon hydrophobic substitutions in a disordered system with non-intuitive effects on folding and function. Aided by sequence-structure analysis, equilibrium, and kinetic spectroscopic studies, we engineer two hydrophobic mutations in the disordered DNA-binding domain of CytR that act synergistically, but not in isolation, to promote structure, compactness, and stability. The double mutant, with properties of a fully ordered domain, exhibits weak cooperativity with a complex and rugged conformational landscape. The mutant, however, binds cognate DNA with an affinity only marginally higher than that of the wild type, though nontrivial differences are observed in the binding to noncognate DNA. Our work provides direct experimental evidence of the dominant role of non-additive hydrophobic effects in shaping the molecular evolution of order in disordered proteins and vice versa, which could be generalized to even folded proteins with implications for protein design and functional manipulation.

Item Type: Journal Article
Additional Information: copyright for this article belongs to AMER CHEMICAL SOC
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Depositing User: Id for Latest eprints
Date Deposited: 13 Mar 2020 10:23
Last Modified: 13 Mar 2020 10:23
URI: http://eprints.iisc.ac.in/id/eprint/62872

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