Identification of ligands for olfactory receptors by functional expression of a receptor library.
The recognition of odorants by olfactory receptors represents the first stage in odor discrimination. Here, we report the generation of an expression library containing a large and diverse repertoire of mouse olfactory receptor sequences in the transmembrane II-VII region. From this library, 80 chimeric receptors were tested against 26 odorants after transfection into HEK-293 cells. Three receptors were identified to respond to micromolecular concentrations of carvone, (-) citronellal, and limonene, respectively. We also found that the mouse I7 receptor, unlike the rat I7 receptor, prefers heptanal instead of octanal, as a result of a single valine-to-isoleucine substitution. This finding represents the beginning of a molecular understanding of odorant recognition. The identification, on a large scale, of cognate receptor-odorant interactions should provide insight into olfactory coding mechanisms.
Krautwurst
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Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Yau
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KW
Reed
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D000109:Acetylcholine / Q000494:pharmacology; D000080462:Acyclic Monoterpenes; D000447:Aldehydes / Q000378:metabolism / Q000494:pharmacology; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D020013:Calcium Signaling; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D015536:Down-Regulation; D005233:Fatty Alcohols / Q000378:metabolism / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D039821:Monoterpenes; D019151:Peptide Library; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000235:genetics / Q000378:metabolism; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013729:Terpenes / Q000378:metabolism / Q000494:pharmacology
D000080462:Acyclic Monoterpenes; D000447:Aldehydes; D005233:Fatty Alcohols; D008024:Ligands; D039821:Monoterpenes; D019151:Peptide Library; D011943:Receptors, Adrenergic, beta; D018035:Receptors, Odorant; D011993:Recombinant Fusion Proteins; D013729:Terpenes; D012243:Rhodopsin; D007545:Isoproterenol; D000109:Acetylcholine; C108217:citronellal