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Intra- versus intermolecular interactions in monellin: Contribution of surface charges to protein assembly

Xue, Wei-Feng LU ; Szczepankiewicz, Olga LU ; Bauer, Mikael LU ; Thulin, Eva LU and Linse, Sara LU (2006) In Journal of Molecular Biology 358(5). p.1244-1255
Abstract
The relative significance of weak non-covalent interactions in biological context has been much debated. Here, we have addressed the contribution of Coulombic interactions to protein stability and assembly experimentally. The sweet protein monellin, a non-covalently linked heterodimeric protein, was chosen for this study because of its ability to spontaneously reconstitute from separated fragments. The reconstitution of monellin mutants containing large surface charge perturbations was compared to the thermostability of structurally equivalent single-chain monellin containing the same sets of mutations under varying salt concentrations. The affinity between monellin fragments is found to correlate with the thermostability of single chain... (More)
The relative significance of weak non-covalent interactions in biological context has been much debated. Here, we have addressed the contribution of Coulombic interactions to protein stability and assembly experimentally. The sweet protein monellin, a non-covalently linked heterodimeric protein, was chosen for this study because of its ability to spontaneously reconstitute from separated fragments. The reconstitution of monellin mutants containing large surface charge perturbations was compared to the thermostability of structurally equivalent single-chain monellin containing the same sets of mutations under varying salt concentrations. The affinity between monellin fragments is found to correlate with the thermostability of single chain monellin, indicating the involvement of the same underlying Coulombic interactions. This confirms that there are no principal differences in the interactions involved in folding and binding. Based on comparison with a previous mutational study involving hydrophobic core residues, the relative contribution of Coulombic interactions to stability and affinity is modest. However, the Coulombic perturbations only affect the association rates of reconstitution in contrast to perturbations involving hydrophobic residues, which affect primarily the dissociation rates. These results indicate that Coulombic interactions are likely to be of main importance for the association of protein assembly, relevant for functions of proteins. (c) 2006 Elsevier Ltd. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
interactions, electrostatic, protein reconstitution, fragment complementation, global analysis, protein folding and binding
in
Journal of Molecular Biology
volume
358
issue
5
pages
1244 - 1255
publisher
Elsevier
external identifiers
  • wos:000237689600006
  • pmid:16574151
  • scopus:33646091307
ISSN
1089-8638
DOI
10.1016/j.jmb.2006.02.069
language
English
LU publication?
yes
id
5a7bd3b2-67d7-402c-a9a7-6bf126a0b3d6 (old id 408601)
date added to LUP
2007-10-21 07:47:43
date last changed
2019-02-27 03:30:51
@article{5a7bd3b2-67d7-402c-a9a7-6bf126a0b3d6,
  abstract     = {The relative significance of weak non-covalent interactions in biological context has been much debated. Here, we have addressed the contribution of Coulombic interactions to protein stability and assembly experimentally. The sweet protein monellin, a non-covalently linked heterodimeric protein, was chosen for this study because of its ability to spontaneously reconstitute from separated fragments. The reconstitution of monellin mutants containing large surface charge perturbations was compared to the thermostability of structurally equivalent single-chain monellin containing the same sets of mutations under varying salt concentrations. The affinity between monellin fragments is found to correlate with the thermostability of single chain monellin, indicating the involvement of the same underlying Coulombic interactions. This confirms that there are no principal differences in the interactions involved in folding and binding. Based on comparison with a previous mutational study involving hydrophobic core residues, the relative contribution of Coulombic interactions to stability and affinity is modest. However, the Coulombic perturbations only affect the association rates of reconstitution in contrast to perturbations involving hydrophobic residues, which affect primarily the dissociation rates. These results indicate that Coulombic interactions are likely to be of main importance for the association of protein assembly, relevant for functions of proteins. (c) 2006 Elsevier Ltd. All rights reserved.},
  author       = {Xue, Wei-Feng and Szczepankiewicz, Olga and Bauer, Mikael and Thulin, Eva and Linse, Sara},
  issn         = {1089-8638},
  keyword      = {interactions,electrostatic,protein reconstitution,fragment complementation,global analysis,protein folding and binding},
  language     = {eng},
  number       = {5},
  pages        = {1244--1255},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Intra- versus intermolecular interactions in monellin: Contribution of surface charges to protein assembly},
  url          = {http://dx.doi.org/10.1016/j.jmb.2006.02.069},
  volume       = {358},
  year         = {2006},
}