期刊
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
卷 13, 期 13, 页码 3112-3120出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpclett.2c00316
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资金
- Science and Engineering Research Board (SERB, Department of Science and Technology, India) [CRG/2019/000084]
- JSPS KAKENHI [JP18H04533, JP19K06577, JP18H02382, JP20K21166]
- Department of Biotechnology (DBT)/Wellcome Trust India Alliance Fellowship [IA/I/18/1/503614]
- Department of Science and Technology (DST)/Science and Engineering Research Board (SERB) Core Research Grant [CRG/2019/003457]
- Indian Institute of Science (IISc)
- Government of India: DST-FIST
- DBT-IISc partnership program
- Council of Scientific and Industrial Research (CSIR)
- UGC-CAS
We investigated the conformational properties of the intrinsically disordered DNA-binding domain of CytR in the presence of polymeric crowder polyethylene glycol (PEG). Our findings demonstrate that the disordered CytR adopts a well-folded conformation in its native ensemble, while the unfolded ensemble collapses and folds with increasing crowder density, regardless of the size of the crowder. We estimate a 10% reduction in the accessible conformational space of a residue in the unfolded state at 300 mg/mL PEG8000. The experimentally constructed PEG-temperature phase diagram shows that entropic effects stabilize disordered CytR by 10 kJ mol-1, promoting the equilibrium towards folded conformations under physiological conditions. Our work highlights the flexible conformational landscape of CytR, reveals the presence of a folded conformation in the disordered ensemble, and proposes a scaling relation for quantifying excluded volume effects on protein stability.
We investigate the conformational properties of the intrinsically disorderedDNA-binding domain of CytR in the presence of the polymeric crowder polyethylene glycol(PEG). Integrating circular dichroism, nuclear magnetic resonance, and single-moleculeFo''rster resonance energy transfer measurements, we demonstrate that disordered CytRpopulates a well-folded minor conformation in its native ensemble, while the unfoldedensemble collapses and folds with an increase in crowder density independent of thecrowder size. Employing a statistical-mechanical model, the effective reduction in theaccessible conformational space of a residue in the unfolded state is estimated to be 10% at300 mg/mL PEG8000, relative to dilute conditions. The experimentally consistent PEG-temperature phase diagram thusconstructed reveals that entropic effects can stabilize disordered CytR by 10 kJ mol-1, driving the equilibrium toward foldedconformations under physiological conditions. Our work highlights the malleable conformational landscape of CytR, the presence ofa folded conformation in the disordered ensemble, and proposes a scaling relation for quantifying excluded volume effects on proteinstability.
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