Structure and stability of recombinant bovine odorant-binding protein: II. Unfolding of the monomeric forms
- Published
- Accepted
- Subject Areas
- Biochemistry, Bioinformatics, Biophysics, Molecular Biology
- Keywords
- odorant-binding protein, ligand binding, disulfide bond, domain swapping, unfolding-refolding reaction, conformational stability
- Copyright
- © 2015 Stepanenko et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ PrePrints) and either DOI or URL of the article must be cited.
- Cite this article
- 2015. Structure and stability of recombinant bovine odorant-binding protein: II. Unfolding of the monomeric forms. PeerJ PrePrints 3:e1452v1 https://doi.org/10.7287/peerj.preprints.1452v1
Abstract
In a family of monomeric odorant-binding proteins (OBPs), bovine OBP (bOBP), that lacks conserved disulfide bond found in other OBPs, occupies unique niche because of its ability to form domain-swapped dimers. In this study, we analyzed conformational stabilities of the recombinant bOBP and its monomeric variants, the bOBP-Gly121+ mutant containing an additional glycine residue after the residue 121 of the bOBP, and the GCC-bOBP mutant obtained from the bOBP-Gly121+ form by introduction of the Trp64Cys/His155Cys double mutation to restore the canonical disulfide bond. We also analyzed the effect of the natural ligand binding on the conformational stabilities of these bOBP variants. Our data are consistent with the conclusion that the unfolding-refolding pathways of the recombinant bOBP and its mutant monomeric forms bOBP-Gly121+ and GCC-bOBP are similar and do not depend on the oligomeric status of the protein. This clearly shows that the information on the unfolding-refolding mechanism is encoded in the structure of the bOBP monomers. However, the process of the bOBP unfolding is significantly complicated by the formation of the domain-swapped dimer, and the rates of the unfolding-refolding reactions essentially depend on the conditions in which the protein is located.
Author Comment
This manuscript is submitted to PeerJ for review. It is a part of series of articlesdedicated to the analysis of the effect of the environmental feature (including the presence of crowding agents) on structural properties and conformational stability of bOBP.