Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments
| Autor: | Olek Maciejak, Jérôme Mathé, Céline Merstorf, Juan Pelta, Philippe Savarin, Bénédicte Thiebot, Patrick A. Curmi, Marie-Jeanne Clément, Manuela Pastoriza-Gallego, Loïc Auvray |
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| Přispěvatelé: | Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Structure et activité des biomolécules normales et pathologiques (SABNP), Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM), Matière et Systèmes Complexes (MSC), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Matière et Systèmes Complexes (MSC (UMR_7057)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE - UMR 8587), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2012 |
| Předmět: |
Protein Folding
Magnetic Resonance Spectroscopy [SDV]Life Sciences [q-bio] Peptides and proteins 010402 general chemistry 01 natural sciences Biochemistry Maltose-Binding Proteins Protein Structure Secondary 03 medical and health sciences Residue (chemistry) Maltose-binding protein chemistry.chemical_compound Nuclear magnetic resonance Guanidine Protein Unfolding 030304 developmental biology Maltose transport 0303 health sciences biology Chemistry Monomers Energy landscape Amides 0104 chemical sciences Folding (chemistry) Crystallography Helix biology.protein Thermodynamics Chemical stability Hydrogen |
| Zdroj: | Biochemistry Biochemistry, 2012, 51 (44), pp.8919-8930. ⟨10.1021/bi3003605⟩ Biochemistry, American Chemical Society, 2012, 51 (44), pp.8919-8930. ⟨10.1021/bi3003605⟩ |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi3003605 |
| Popis: | International audience; Being able to differentiate local fluctuations from global folding−unfolding dynamics of a protein is of major interest for improving our understanding of structure−function determinants. The maltose binding protein (MBP), a protein that belongs to the maltose transport system, has a structure composed of two globular domains separated by a rigid-body “hinge bending”. Here we determined, by using hydrogen exchange (HX) nuclear magnetic resonance experiments, the apparent stabilization free energies of 101 residues of MBP bound to β-cyclodextrin (MBP−βCD) under native conditions. We observed that the last helix of MBP (helix α14) has a lower protection factor than the rest of the protein. Further, HX experiments were performed using guanidine hydrochloride under subdenaturing conditions to discriminate between local fluctuations and global unfolding events and to determine the MBP−βCD energy landscape. The results show that helix α4 and a part of helices α5 and α6 are clearly grouped into a subdenaturing folding unit and represent a partially folded intermediate under native conditions. In addition, we observed that amide protons located in the hinge between the two globular domains share similar ΔGgu app and m values and should unfold simultaneously. These observations provide new points of view for improving our understanding of the thermodynamic stability and the mechanisms that drive folding−unfolding dynamics of proteins. |
| Databáze: | OpenAIRE |
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