PUBLICATION
Full rescue of an inactive olfactory receptor mutant by elimination of an allosteric ligand-gating site
- Authors
- Sharma, K., Balfanz, S., Baumann, A., Korsching, S.
- ID
- ZDB-PUB-180627-6
- Date
- 2018
- Source
- Scientific Reports 8: 9631 (Journal)
- Registered Authors
- Korsching, Sigrun
- Keywords
- none
- MeSH Terms
-
- Allosteric Site
- Binding Sites
- Cadaverine/metabolism
- Ligands
- Models, Molecular
- Mutation*
- Protein Conformation
- Receptors, Odorant/chemistry*
- Receptors, Odorant/genetics
- Receptors, Odorant/metabolism*
- PubMed
- 29941999 Full text @ Sci. Rep.
Citation
Sharma, K., Balfanz, S., Baumann, A., Korsching, S. (2018) Full rescue of an inactive olfactory receptor mutant by elimination of an allosteric ligand-gating site. Scientific Reports. 8:9631.
Abstract
Ligand-gating has recently been proposed as a novel mechanism to regulate olfactory receptor sensitivity. TAAR13c, the zebrafish olfactory receptor activated by the death-associated odor cadaverine, appears to possess an allosteric binding site for cadaverine, which was assumed to block progress of the ligand towards the internal orthosteric binding-and-activation site. Here we have challenged the suggested gating mechanism by modeling the entry tunnel for the ligand as well as the ligand path inside the receptor. We report an entry tunnel, whose opening is blocked by occupation of the external binding site by cadaverine, confirming the hypothesized gating mechanism. A multistep docking algorithm suggested a plausible path for cadaverine from the allosteric to the orthosteric binding-and-activation site. Furthermore we have combined a gain-of-function gating site mutation and a loss-of-function internal binding site mutation in one recombinant receptor. This receptor had almost wildtype ligand affinities, consistent with modeling results that showed localized effects for each mutation. A novel mutation of the suggested gating site resulted in increased receptor ligand affinity. In summary both the experimental and the modeling results provide further evidence for the proposed gating mechanism, which surprisingly exhibits pronounced similarity to processes described for some metabotropic neurotransmitter receptors.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping