Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Biol Chem
2017 Nov 17;29246:18916-18923. doi: 10.1074/jbc.M117.810374.
Show Gene links
Show Anatomy links
Mutant cycle analysis identifies a ligand interaction site in an odorant receptor of the malaria vector Anopheles gambiae.
Rahman S, Luetje CW.
???displayArticle.abstract???
Lack of information about the structure of insect odorant receptors (ORs) hinders the development of more effective repellants to control disease-transmitting insects. Mutagenesis and functional analyses using agonists to map the odorant-binding sites of these receptors have been limited because mutations distant from an agonist-binding site can alter agonist sensitivity. Here we use mutant cycle analysis, an approach for exploring the energetics of protein-protein or protein-ligand interactions, with inhibitors, to identify a component of the odorant-binding site of an OR from the malaria vector, Anopheles gambiae The closely related odorant-specificity subunits Agam/Or15 and Agam/Or13 were each co-expressed with Agam/Orco (odorant receptor co-receptor subunit) in Xenopus oocytes and assayed by two-electrode voltage clamp electrophysiology. We identified (-)-fenchone as a competitive inhibitor with different potencies at the two receptors and used this difference to screen a panel of 37 Agam/Or15 mutants, surveying all positions that differ between Agam/Or15 and Agam/Or13 in the transmembrane and extracellular regions, identifying position 195 as a determinant of (-)-fenchone sensitivity. Inhibition by (-)-fenchone and six structurally related inhibitors of Agam/Or15 receptors containing each of four different hydrophobic residues at position 195 served as input data for mutant cycle analysis. Several mutant cycles, calculated from the inhibition of two receptors by each of two ligands, yielded coupling energies of ≥1 kcal/mol, indicating a close, physical interaction between the ligand and residue 195 of Agam/Or15. This approach should be useful in further expanding our knowledge of odorant-binding site structures in ORs of disease vector insects.
Benton,
Atypical membrane topology and heteromeric function of Drosophila odorant receptors in vivo.
2006, Pubmed
Benton,
Atypical membrane topology and heteromeric function of Drosophila odorant receptors in vivo.
2006,
Pubmed Carey,
Odorant reception in the malaria mosquito Anopheles gambiae.
2010,
Pubmed Carraher,
Towards an understanding of the structural basis for insect olfaction by odorant receptors.
2015,
Pubmed Carter,
The use of double mutants to detect structural changes in the active site of the tyrosyl-tRNA synthetase (Bacillus stearothermophilus).
1984,
Pubmed Colquhoun,
Binding, gating, affinity and efficacy: the interpretation of structure-activity relationships for agonists and of the effects of mutating receptors.
1998,
Pubmed DeGennaro,
orco mutant mosquitoes lose strong preference for humans and are not repelled by volatile DEET.
2013,
Pubmed Erdelyan,
Functional validation of the carbon dioxide receptor genes in Aedes aegypti mosquitoes using RNA interference.
2012,
Pubmed Gibson,
Visual and olfactory responses of haematophagous Diptera to host stimuli.
1999,
Pubmed Guidobaldi,
Morphology and physiology of the olfactory system of blood-feeding insects.
2014,
Pubmed Hallem,
Olfaction: mosquito receptor for human-sweat odorant.
2004,
Pubmed Hallem,
Coding of odors by a receptor repertoire.
2006,
Pubmed Hallem,
The spatial code for odors is changed by conditioning.
2004,
Pubmed Hidalgo,
Revealing the architecture of a K+ channel pore through mutant cycles with a peptide inhibitor.
1995,
Pubmed
,
Xenbase Horovitz,
Strategy for analysing the co-operativity of intramolecular interactions in peptides and proteins.
1990,
Pubmed Hughes,
A determinant of odorant specificity is located at the extracellular loop 2-transmembrane domain 4 interface of an Anopheles gambiae odorant receptor subunit.
2014,
Pubmed
,
Xenbase Jones,
Functional conservation of an insect odorant receptor gene across 250 million years of evolution.
2005,
Pubmed Krieger,
A candidate olfactory receptor subtype highly conserved across different insect orders.
2003,
Pubmed Larsson,
Or83b encodes a broadly expressed odorant receptor essential for Drosophila olfaction.
2004,
Pubmed Leff,
Further concerns over Cheng-Prusoff analysis.
1993,
Pubmed Lu,
Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae.
2007,
Pubmed McMeniman,
Multimodal integration of carbon dioxide and other sensory cues drives mosquito attraction to humans.
2014,
Pubmed Nakagawa,
Controversy and consensus: noncanonical signaling mechanisms in the insect olfactory system.
2009,
Pubmed Neuhaus,
Odorant receptor heterodimerization in the olfactory system of Drosophila melanogaster.
2005,
Pubmed Nichols,
Transmembrane segment 3 of Drosophila melanogaster odorant receptor subunit 85b contributes to ligand-receptor interactions.
2010,
Pubmed
,
Xenbase Nyce,
Mapping spatial relationships between residues in the ligand-binding domain of the 5-HT3 receptor using a molecular ruler.
2010,
Pubmed Pask,
Heteromeric Anopheline odorant receptors exhibit distinct channel properties.
2011,
Pubmed Pellegrino,
A natural polymorphism alters odour and DEET sensitivity in an insect odorant receptor.
2011,
Pubmed Penzotti,
Specific neosaxitoxin interactions with the Na+ channel outer vestibule determined by mutant cycle analysis.
2001,
Pubmed
,
Xenbase Pitts,
A highly conserved candidate chemoreceptor expressed in both olfactory and gustatory tissues in the malaria vector Anopheles gambiae.
2004,
Pubmed Ranganathan,
Spatial localization of the K+ channel selectivity filter by mutant cycle-based structure analysis.
1996,
Pubmed Schreiber,
Energetics of protein-protein interactions: analysis of the barnase-barstar interface by single mutations and double mutant cycles.
1995,
Pubmed Smallegange,
Sweaty skin: an invitation to bite?
2011,
Pubmed Spyropoulos,
TMRPres2D: high quality visual representation of transmembrane protein models.
2004,
Pubmed Thomas-Tran,
Mutant cycle analysis with modified saxitoxins reveals specific interactions critical to attaining high-affinity inhibition of hNaV1.7.
2016,
Pubmed Tondi,
Targeting class A and C serine β-lactamases with a broad-spectrum boronic acid derivative.
2014,
Pubmed Vosshall,
A unified nomenclature system for the insect olfactory coreceptor.
2011,
Pubmed Wang,
Molecular basis of odor coding in the malaria vector mosquito Anopheles gambiae.
2010,
Pubmed
,
Xenbase Wicher,
dOr83b--receptor or ion channel?
2009,
Pubmed Wicher,
Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels.
2008,
Pubmed Willcockson,
Orientation of d-tubocurarine in the muscle nicotinic acetylcholine receptor-binding site.
2002,
Pubmed Zwiebel,
Olfactory regulation of mosquito-host interactions.
2004,
Pubmed
