Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments - Archive ouverte HAL Access content directly
Journal Articles Biochemistry Year : 2012

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

(1) , (1, 2) , (1) , (1) , (1) , (2) , (1) , (3) , (2) , (2)
1
2
3

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.
Not file

Dates and versions

hal-02159344 , version 1 (18-06-2019)

Identifiers

Cite

Céline Merstorf, Olek Maciejak, Jérôme Mathé, Manuela Pastoriza-Gallego, Bénédicte Thiebot, et al.. Mapping the Conformational Stability of Maltose Binding Protein at the Residue Scale Using Nuclear Magnetic Resonance Hydrogen Exchange Experiments. Biochemistry, 2012, 51 (44), pp.8919-8930. ⟨10.1021/bi3003605⟩. ⟨hal-02159344⟩
35 View
0 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More