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Activation Barrier-Limited Folding and Conformational Sampling of a Dynamic Protein Domain
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology. Stockholm Univ, Dept Biochem & Biophys, SE-10691 Stockholm, Sweden..
Univ Cambridge, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England..
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
Univ Rome, Inst Pasteur, Fdn Cenci Bolognetti, I-00185 Rome, Italy.;Univ Rome, Ist Biol & Patol Mol, CNR, Dipartimento Sci Biochim A Rossi Fanelli Sapienza, I-00185 Rome, Italy.;Univ Cambridge, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England..
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2016 (English)In: Biochemistry, ISSN 0006-2960, E-ISSN 1520-4995, Vol. 55, no 37, 5289-5295 p.Article in journal (Refereed) Published
Abstract [en]

Folding reaction mechanisms of globular protein domains have been extensively studied by both experiment and simulation and found to be highly concerted chemical reactions in which numerous noncovalent bonds form in an apparent two-state fashion. However, less is known regarding intrinsically disordered proteins because their folding can usually be studied only in conjunction with binding to a ligand. We have investigated by kinetics the folding mechanism of such a disordered protein domain, the nuclear coactivator-binding domain (NCBD) from CREB-binding protein. While a previous computational study suggested that NCBD folds without an activation free energy barrier, our experimental data. demonstrate that NCBD, despite its highly dynamic structure, displays relatively slow folding (similar to 10 ms at 277 K) consistent with a barrier-limited process. Furthermore, the folding kinetics corroborate previous nuclear magnetic resonance data showing that NCBD exists in two folded conformations and one more denatured conformation at equilibrium and, thus, that the folding mechanism is a three-state mechanism. The refolding kinetics is limited by unfolding of the less populated folded conformation, suggesting that the major route for interconversion between the two folded states is via the denatured State. Because the two folded conformations have been suggested to bind distinct ligands, our results have mechanistic implications for conformational sampling in protein protein interactions.

Place, publisher, year, edition, pages
2016. Vol. 55, no 37, 5289-5295 p.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
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URN: urn:nbn:se:uu:diva-306263DOI: 10.1021/acs.biochem.6b00573ISI: 000384038700010PubMedID: 27542287OAI: oai:DiVA.org:uu-306263DiVA: diva2:1040200
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Swedish Research Council
Available from: 2016-10-26 Created: 2016-10-26 Last updated: 2016-10-26Bibliographically approved

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Andersson, EvaJemth, Per
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