Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments

Céline Merstorf; Olek Maciejak; Jérôme Mathé; Manuela Pastoriza-Gallego; Bénédicte Thiebot; Marie-Jeanne Clément; Juan Pelta; Loïc Auvray; Patrick Curmi; Philippe Savarin

Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments

Céline Merstorf ¹ Olek Maciejak ^{1, 2} Jérôme Mathé ¹ Manuela Pastoriza-Gallego ¹ Bénédicte Thiebot ¹ Marie-Jeanne Clément ² Juan Pelta ¹ Loïc Auvray ³ Patrick Curmi ² Philippe Savarin ²

1 LAMBE - UMR 8587 - Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement

2 SABNP - Structure et activité des biomolécules normales et pathologiques

3 MSC (UMR_7057) - Matière et Systèmes Complexes

Abstract : 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.

Document type :

Journal articles

Domain :

Life Sciences [q-bio]

Complete list of metadatas

https://hal-univ-evry.archives-ouvertes.fr/hal-02159344
Contributor : Olek Maciejak <>
Submitted on : Tuesday, June 18, 2019 - 4:34:49 PM
Last modification on : Friday, October 25, 2019 - 11:02:17 AM

Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments

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Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments

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