Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes.
Here, we provide the first evidence for functional expression of a human olfactory receptor protein (OR17-40) and show that recombinant olfactory receptors can be functionally expressed in heterologous systems. A mixture of 100 different odorants (Henkel 100) elicited a transient increase in intracellular [Ca(2+)] in human embryonic kidney 293 (HEK293) cells stably or transiently transfected with the plasmid pOR17-40. By subdividing the odorant mixture into progressively smaller groups, we identified a single component that represented the only effective substance: helional. Only the structurally closely related molecule heliotroplyacetone also activated the receptor. Other compounds, including piperonal, safrole, and vanillin, were completely ineffective. Mock-transfected cells and cells transfected with other receptors showed no change in intracellular [Ca(2+)] in response to odor stimulation. We were also able to functionally express OR17-40 in Xenopus laevis oocytes. Coexpression of a "reporter" channel allowed measurement of the response of oocytes injected with the cRNA of the human receptor to the odor mixture Henkel 100. The effective substances were the same (helional, heliotropylacetone) as those identified by functionally expressing the receptor in HEK293 cells and were active at the same, lower micromolar concentration. These findings open the possibility of now characterizing the sensitivity and specificity of many, if not all, of the hundreds of different human olfactory receptors.
Wetzel
C H
CH
Lehrstuhl für Zellphysiologie, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Oles
M
M
Wellerdieck
C
C
Kuczkowiak
M
M
Gisselmann
G
G
Hatt
H
H
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D018076:DNA, Complementary; D005260:Female; D006801:Humans; D007668:Kidney; D008564:Membrane Potentials / Q000187:drug effects; D008959:Models, Neurological; D008969:Molecular Sequence Data; D009812:Odorants; D009865:Oocytes / Q000502:physiology; D017433:Protein Structure, Secondary; D018075:RNA, Complementary / Q000378:metabolism; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection; D014982:Xenopus laevis
D018076:DNA, Complementary; D018075:RNA, Complementary; D018035:Receptors, Odorant; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D002118:Calcium