Show simple item record Chang, YC Oas, TG
dc.coverage.spatial United States 2011-06-21T17:22:09Z 2010-06-29
dc.identifier.citation Biochemistry, 2010, 49 (25), pp. 5086 - 5096
dc.description.abstract Understanding the interconversion between thermodynamically distinguishable states present in a protein folding pathway provides not only the kinetics and energetics of protein folding but also insights into the functional roles of these states in biological systems. The protein component of the bacterial RNase P holoenzyme from Bacillus subtilis (P protein) was previously shown to be unfolded in the absence of its cognate RNA or other anionic ligands. P protein was used in this study as a model system to explore general features of intrinsically disordered protein (IDP) folding mechanisms. The use of trimethylamine N-oxide (TMAO), an osmolyte that stabilizes the unliganded folded form of the protein, enabled us to study the folding process of P protein in the absence of ligand. Transient stopped-flow kinetic traces at various final TMAO concentrations exhibited multiphasic kinetics. Equilibrium "cotitration" experiments were performed using both TMAO and urea during the titration to produce a urea-TMAO titration surface of P protein. Both kinetic and equilibrium studies show evidence of a previously undetected intermediate state in the P protein folding process. The intermediate state is significantly populated, and the folding rate constants are relatively slow compared to those of intrinsically folded proteins similar in size and topology. The experiments and analysis described serve as a useful example for mechanistic folding studies of other IDPs.
dc.format.extent 5086 - 5096
dc.language eng
dc.language.iso en_US en_US
dc.relation.ispartof Biochemistry
dc.relation.isversionof 10.1021/bi100222h
dc.subject Bacillus subtilis
dc.subject Bacterial Proteins
dc.subject Circular Dichroism
dc.subject Kinetics
dc.subject Ligands
dc.subject Methylamines
dc.subject Protein Folding
dc.subject Spectrometry, Fluorescence
dc.subject Thermodynamics
dc.subject Tryptophan
dc.title Osmolyte-induced folding of an intrinsically disordered protein: folding mechanism in the absence of ligand.
dc.title.alternative en_US
dc.type Journal Article
dc.description.version Version of Record en_US 2010-6-29 en_US
duke.description.endpage 5096 en_US
duke.description.issue 25 en_US
duke.description.startpage 5086 en_US
duke.description.volume 49 en_US
dc.relation.journal Biochemistry en_US
pubs.issue 25
pubs.organisational-group /Duke
pubs.organisational-group /Duke/School of Medicine
pubs.organisational-group /Duke/School of Medicine/Basic Science Departments
pubs.organisational-group /Duke/School of Medicine/Basic Science Departments/Biochemistry
pubs.organisational-group /Duke/School of Medicine/Institutes and Centers
pubs.organisational-group /Duke/School of Medicine/Institutes and Centers/Duke Cancer Institute
pubs.publication-status Published
pubs.volume 49
dc.identifier.eissn 1520-4995

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