Skip to main content
NCBI home page
Scientific Data logoLink to Scientific Data
. 2015 Feb 3;2:150002. doi: 10.1038/sdata.2015.2

Human olfactory receptor responses to odorants

Joel D Mainland 1,2,3,a, Yun R Li 2, Ting Zhou 2, Wen Ling L Liu 2, Hiroaki Matsunami 2,4
PMCID: PMC4412152  PMID: 25977809

Abstract

Although the human olfactory system is capable of discriminating a vast number of odors, we do not currently understand what chemical features are encoded by olfactory receptors. In large part this is due to a paucity of data in a search space covering the interactions of hundreds of receptors with billions of odorous molecules. Of the approximately 400 intact human odorant receptors, only 10% have a published ligand. Here we used a heterologous luciferase assay to screen 73 odorants against a clone library of 511 human olfactory receptors. This dataset will allow other researchers to interrogate the combinatorial nature of olfactory coding.

Subject terms: Behavioural genetics, Olfactory receptors

Background & Summary

Previous functional analysis of olfactory receptors (ORs) in olfactory neurons and in heterologous cells found that different odorants are recognized by unique, but overlapping ensembles of ORs1–4. These findings suggest that specific patterns of ORs activated by an odorant code for the odorant’s identity, but there are few, if any, explicit predictions relating OR activity patterns to olfactory perception.

Matching mammalian ORs to ligands has seen limited success, and the picture is even worse when considering human ORs; ligands have been published for only 49 of the approximately 400 intact human ORs5–21. This lack of data is a critical bottleneck in the field; matching ligands to ORs is critical for understanding the olfactory system at all levels and is essential for building viable models of olfaction. The characterization of OR responses to ligands in the empty neuron system of Drosophila melanogaster 22 has allowed researchers in the field to choose rationally diverse odorant sets23 and specifically manipulate subpopulations of ORs to dissect olfactory coding24,25. Extending this idea by matching odorants to human ORs has the added advantage that humans can directly communicate their perception of odor intensity, pleasantness, and quality.

In addition, understanding the role of a single OR in olfactory perception allows us to look at evolutionary changes in OR genes in a new light. For example, the knowledge that Tas1r2 is a pseudogene in seven of twelve species in the order Carnivora 26 is difficult to interpret in isolation. The knowledge that Tas1r2 is the primary mediator of sweet taste in mice, however, suggests that carnivores do not need to taste sweet and therefore there is no selective pressure on the gene. As several genome sequencing projects are examining both genetic variation within humans27 and across species28, understanding the role of OR genes in olfactory perception becomes crucial to the interpretation of how and why genetic changes occur over the course of evolution.

In a recent manuscript we conducted a high-throughput screen of 511 human odorant receptors against 73 odorants15. The resulting screen identified agonists for 27 odorant receptors, including 18 that were previously orphan receptors. We went on to characterize how genetic variation in these receptors alters both in vitro responses and influences olfactory perception. In this manuscript we present the full screening data to permit wider reuse and reanalysis.

In summary, this dataset addresses a major bottleneck in the field, namely how the physical stimulus in olfaction is transduced into receptor responses. In addition, the G-protein coupled receptor class accounts for approximately 50% of therapeutic drug targets29. The ORs, being GPCRs, offer the opportunity to examine the strategies employed by this receptor class to recognize a wide variety of ligand structural features and thus will provide insight into fundamental principles of ligand recognition by GPCRs. Matching odorants to ORs will provide a valuable resource to the field and allow more specific explorations of links between odor, behavior and ecology.

Methods

These methods are expanded from descriptions in our previous work15.

Cloning

OR open reading frames were amplified from genomic DNA using Phusion polymerase and subcloned into pCI expression vectors (Promega) containing the first 20 residues of human rhodopsin (Rho tag). Human ORs were amplified from the pooled genomic DNA of 20 participants from the International Hapmap Consortium, while mouse ORs were amplified from the genomic DNA of C57/BL6 mice. The sequences of the cloned receptors were verified by sequencing (3100 Genetic Analyzer, Applied Biosystems). Clones that were present in the 1000 Genomes Project, but not cloned from our pooled genomic DNA sample, were created using an overlap extension polymerase chain reaction protocol30.

Luciferase assay

The Dual-Glo Luciferase Assay System (Promega) was used to measure receptor responses as previously described31. Hana3A cells were transfected with 5 ng/well of RTP1S32, 5 ng/well of pRL-SV40, 10 ng/well of CRE-luciferase, 2.5 ng/well of M3 (ref. 33), and 5 ng/well of odorant receptor. 1 M odorant stocks are diluted in DMSO. 24 hours following transfection, transfection media was removed and replaced with the appropriate concentration of odor diluted from the 1 M stocks in CD293 (Gibco). Four hours following odor stimulation luminescence was measured using a Polarstar Optima plate reader (BMG). All luminescence values were divided by the Renilla Luciferase activity to control for transfection efficiency in a given well. Data were analyzed with Microsoft Excel, GraphPad Prism 4, and MATLAB.

Primary screen design

Our screen design is outlined in Figure 1. In the primary screen we stimulated 511 human ORs with 73 odorants used in previous psychophysical testing12,34. We applied the majority of odorants at a concentration of 100 μM. All plates in the primary screen included 85 test wells, five broadly-tuned odorant receptors (Olfr1079, OR2W1, Olfr1377, Olfr73, Olfr1341), and six wells transfected with Oflr544 which served as a standard. Of the six wells transfected with Olfr544, three were challenged with the diluent (CD293) and three were challenged with 10 μM of a known ligand for Olfr544 (nonanedioic acid). Each screening run consisted of twelve plates where each of the 96-wells were transfected with the same set of receptors. One plate had no odor in all test wells and served as a baseline. The other eleven plates were each challenged with a different test odor.

Figure 1. Outline of the screening procedure.

Figure 1

Open in a new tab

This figure was reprinted from our previous publication15, where it was included as Supplementary Figure 1.

Secondary screen design

To rank hits from the Primary Screen we standardized each plate, setting the mean Olfr544 response to nonanedioic acid minus the mean Olfr544 response to the no-odor control to a value of 1. We then subtracted the baseline response for each receptor from the no-odor plate from the response to the odor challenge and ranked the resulting values. We selected the top 5% of odorant/receptor pairs from the primary screen, although not more than the top ten ligands for a given receptor. We then performed a secondary screen in which each odorant receptor was tested against a no-odor control as well as 1, 10 and 100 μM of odor. Each comparison was performed in triplicate, where each measure was collected from separate wells, but each well contained cells from the same parent plate of cells. Note that we began the secondary screen before completion of the entire primary screen, so some odor/receptor combinations outside of the overall top 5% were tested.

Dose-response design

We then constructed dose-response curves using concentrations ranging from 10 nM to 10 mM for the odor/receptor pairs that were significantly different from baseline in the Secondary Screen. Each odorant receptor-odorant dose was tested in triplicate, where each measure was collected from separate wells, but each well contains cells from the same parent plate of cells. A vector-only control was included for each odorant. We fit the data to a sigmoidal curve. We counted an odorant as an agonist if the 95% confidence intervals of the top and bottom parameters did not overlap, the standard deviation of the fitted log EC50 was less than 1 log unit, and the extra sums-of-squares test confirmed that the odorant activated the receptor significantly more than the control, which was transfected with an empty vector. This data identified 25 odorant receptors with a significant response to at least one agonist15 (Figures 2 and 3).

Figure 2. Normalized dose-response curves of the receptor encoded by the most common functional allele for 25 receptors.

Figure 2

Open in a new tab

The responses of cells transfected with either a plasmid encoding the indicated odorant receptor or an empty vector to the indicated odorants. Responses have been normalized such that each receptor has a minimum response of zero and a maximum response of one. Error bars, s.e.m. over three replicates. Abbreviations for the odorants are as follows: +CAR are shown, (+)-carvone; LIN, linalool; GA, geranyl acetate; COUM, coumarin; OTHI, octanethiol; C3HEX, cis-3-hexen-1-ol; EUG, eugenol; EUGME, eugenol methyl ether; ANIS, anisaldehyde; ANDI, 4,16-androstadien-3-one; AND, 5α-androst-16-en-3-one; DMHDMF, caramel furanone; +MEN, (+)-menthol; 3PPP, 3-phenyl propyl propionate; VAN, vanillin; LYR, lyral; 2EF, 2-ethyl fenchol; IVA, isovaleric acid; APA, allyl phenyl acetate. This figure was modified from our previous publication15, where it was included as Figure 1.

Figure 3. Dose-response curves of the receptor encoded by the most common functional allele for 25 receptors.

Figure 3

Open in a new tab

The responses of cells transfected with either a plasmid encoding the indicated odorant receptor or an empty vector to the indicated odorants. Error bars, s.e.m. over three replicates. Abbreviations for the odorants are as follows: +CAR are shown, (+)-carvone; LIN, linalool; GA, geranyl acetate; COUM, coumarin; OTHI, octanethiol; C3HEX, cis-3-hexen-1-ol; EUG, eugenol; EUGME, eugenol methyl ether; ANIS, anisaldehyde; ANDI, 4,16-androstadien-3-one; AND, 5α-androst-16-en-3-one; DMHDMF, caramel furanone; +MEN, (+)-menthol; 3PPP, 3-phenyl propyl propionate; VAN, vanillin; LYR, lyral; 2EF, 2-ethyl fenchol; IVA, isovaleric acid; APA, allyl phenyl acetate. This figure was modified from our previous publication15, where it was included as Figure 1.

Data Records

The data for this manuscript have been deposited in figshare (Data Citation 1). A summary of the clones tested in each phase of the screen is presented in Supplementary Table 1.

Data record 1—primary screen

The raw screening results are presented in a tab-separated values file (Data Citation 1). Each row represents an experiment from a single well.

Plate. A unique ID for a 96-well plate on a given date.

Well. A number assigned to each well of the 96-well plate. The wells are sequentially numbered with the upper-leftmost well assigned as 1 and the lower-leftmost well assigned as 85 (see Figure 4).

Figure 4. Plate layout for the primary screen.

Figure 4

Open in a new tab

Screens were set up with a master transfection plate for each day. The master transfection plate was used to transfect twelve plates. Each plate was then stimulated with a different odor. Eleven wells were reserved for broadly tuned receptors and a standard to validate the protocol.

Concentration. The concentration of the odorant applied in uM. A ‘9999’ indicates no odor was applied (DMSO was diluted 1:10,000 in CD293).

Luc. The number of photons counted by the plate reader when the well was treated with the luciferase substrate. This is the cAMP reporter, and therefore correlates with receptor responses to odorants.

RL. The number of photons counted by the plate reader when the well was treated with the Renilla luciferase substrate. This is the constitutively active reporter, which serves as a control for cell death and transfection efficiency.

OR. A unique ID for each olfactory receptor clone.

Odor. A unique ID for the odorant applied to the well.

Date. The date the experiment was run in MM/DD/YY format.

Data record 2—secondary screen

The raw screening results are presented in a tab-separated values file (Data Citation 1). Each row represents an experiment from a single well.

Date. The date the experiment was run in MM/DD/YY format.

OR. A unique ID for each olfactory receptor clone.

Odor. A unique ID for the odorant applied to the well.

Concentration. The concentration of the odorant applied in uM. A ‘0’ indicates no odor was applied (CD293 only). Note that rows for the ‘no odor’ condition will contain an ‘Odor’ label to facilitate pairing controls with the matched experiments at other concentrations.

NormalizedLuc. The Luc/RL ratio from each well.

Data record 3—dose-response

The Luc/RL ratios are presented in a tab-separated values file (Data Citation 1). Each row represents an experiment from a single well. The EC50 for each odor/receptor pair that passed this phase of screening is listed in Table 1 (available online only).

Table 1. Log EC50s for all OR/odor pairs that pass the three statistical criteria outlined in the methods.

OR Odor EC50 OdorName Gene Accession
1030 1341 −5 sandalwood OR14A16 Indel KP290123
1034 1101 −4 anisaldehyde OR6P1 KP290127
1034 1337 −5 phenyl acetaldehyde OR6P1 KP290127
1037 1379 −3 lyral OR10J5 KP290130
1037 1379 −5 lyral OR10J5 KP290130
1042 1285 −3 2-decenal OR14A2 KP290135
1044 1115 −6 coumarin OR1C1 KP290137
1044 1328 −6 linalool OR1C1 KP290137
1062 1286 −5 2-ethylfenchol OR11A1 KP290155
1067 1300 −5 cis-3-hexen-1-ol OR2W1 KP290160
1067 1300 −5 cis-3-hexen-1-ol OR2W1 KP290160
1073 1307 −4 ethyl vanillin OR2J2 T111A KP290166
1073 1290 −5 androstenone OR2J2 T111A KP290166
1073 1416 −5 butyl anthranilate OR2J2 T111A KP290166
1073 1311 −6 eugenol methyl ether OR2J2 T111A KP290166
1073 1337 −6 phenyl acetaldehyde OR2J2 T111A KP290166
1073 1324 −5 isoeugenol OR2J2 T111A KP290166
1073 1300 −5 cis-3-hexen-1-ol OR2J2 T111A KP290166
1073 1290 −3 androstenone OR2J2 T111A KP290166
1082 1310 −3 eugenol acetate OR2F1 KP290175
1105 1328 −4 linalool OR1N2 W23R/V230G/T287M KP290194
1111 1111 −5 cinnamaldehyde OR9G1 KP290198
1115 1115 −5 coumarin OR5P3 KP290202
1119 1287 −6 2-methoxy-4-methylphenol OR10G4 A9V/M134V/V195E/R235G/K295Q KP290206
1119 1346 −4 vanillin OR10G4 A9V/M134V/V195E/R235G/K295Q KP290206
1119 1346 −3 vanillin OR10G4 A9V/M134V/V195E/R235G/K295Q KP290206
1120 1309 −8 eugenol OR10G7 KP290207
1129 1325 −4 isovaleric acid OR51E1 KP290215
1129 1325 −3 isovaleric acid OR51E1 KP290215
1129 1325 −3 isovaleric acid OR51E1 KP290215
1129 1325 −5 isovaleric acid OR51E1 KP290215
1134 1326 −4 jasmine OR4A4 KP290220
1136 1326 −4 jasmine OR1S2 KP290222
1137 1326 −4 jasmine OR1S2 I46T KP290223
1142 1341 −4 sandalwood OR8D1 KP290227
1142 1396 −5 caramel furanone OR8D1 KP290227
1155 1325 −3 isovaleric acid OR4C12 V283L KP290235
1161 1290 −3 androstenone OR4F17 KP290239
1183 1191 −7 allyl phenylacetate OR51L1 KP290258
1193 1326 −5 jasmine OR4X2 KP290267
1195 1282 −7 (+)-menthol OR8K3 L122R KP290269
1195 1281 −6 (−)-menthol OR8K3 L122R KP290269
1195 1282 −6 (+)-menthol OR8K3 L122R KP290269
1205 1069 −3 propionic acid OR51E2 KP290277
1206 1295 −4 butyric acid OR51D1 KP290278
1219 1324 −4 isoeugenol OR10A6 KP290290
1230 1295 −4 butyric acid OR5D14 KP290299
1251 1299 −4 cinnamon OR10AG1 KP290316
1257 1325 −4 isovaleric acid OR1E1 KP290321
1263 1325 −4 isovaleric acid OR11H4 KP290326
1265 1316 −4 geranyl acetate OR4L1 R52S KP290327
1265 1316 −4 geranyl acetate OR4L1 R52S KP290327
1272 1346 −3 vanillin OR10G3 S73G KP290332
1272 1346 −3 vanillin OR10G3 S73G KP290332
1275 1325 −4 isovaleric acid OR11H6 KP290335
1277 1326 −3 jasmine OR4N4 KP290337
1282 1274 −5 1-octanethiol OR2C1 C149W KP290341
1282 1203 −6 thioglycolic acid OR2C1 C149W KP290341
1282 1337 −5 phenyl acetaldehyde OR2C1 C149W KP290341
1285 1310 −5 eugenol acetate OR4D2 C97S/L187F KP290344
1288 1190 −3 dihydrojasmone OR3A1 KP290347
1288 1299 −4 cinnamon OR3A1 KP290347
1291 1341 −5 sandalwood OR10H1 KP290349
1298 1290 −5 androstenone OR7D4 KP290356
1298 1290 −6 androstenone OR7D4 KP290356
1298 1290 −6 androstenone OR7D4 KP290356
1298 1290 −5 androstenone OR7D4 KP290356
1298 1290 −5 androstenone OR7D4 KP290356
1299 1315 −5 androstadienone OR7C1 KP290357
1299 1315 −5 androstadienone OR7C1 KP290357
1299 1290 −4 androstenone OR7C1 KP290357
1306 1309 −4 eugenol OR52B6 T36A/L90H/A146T/H149R/V267I KP290361
1339 1310 −4 eugenol acetate OR13C3 G2C KP290384
1350 1328 −4 linalool OR1N2 W37R/V244G/T301M KP290391
1351 1295 −4 butyric acid OR2T1 KP290392
1362 1295 −5 butyric acid OR5AL1 KP290394
1368 1281 −5 (−)-menthol OR10X1 KP290399
1376 1316 −4 geranyl acetate OR1D2 KP290405
1377 1081 −3 geraniol OR2M7 KP290406
1378 1081 −3 geraniol OR2M7 F35L/V78A/C178F KP290407
1387 1316 −6 geranyl acetate OR2A25 KP290416
1387 1360 −6 quinoline OR2A25 KP290416
1387 1316 −5 geranyl acetate OR2A25 KP290416
1402 1307 −4 ethyl vanillin OR2J2 Y74H/T111A/V146A/T218A KP290431
1402 1337 −5 phenyl acetaldehyde OR2J2 Y74H/T111A/V146A/T218A KP290431
1402 1324 −4 isoeugenol OR2J2 Y74H/T111A/V146A/T218A KP290431
1403 1324 −4 isoeugenol OR2J2 KP290432
1403 1337 −4 phenyl acetaldehyde OR2J2 KP290432
1403 1307 −4 ethyl vanillin OR2J2 KP290432
1404 1300 −5 cis-3-hexen-1-ol OR2J2 KP290433
1404 1311 −7 eugenol methyl ether OR2J2 KP290433
1404 1324 −5 isoeugenol OR2J2 KP290433
1404 1337 −6 phenyl acetaldehyde OR2J2 KP290433
1406 1111 −4 cinnamaldehyde OR2C1 KP290435
1406 1274 −8 1-octanethiol OR2C1 KP290435
1407 1274 −6 1-octanethiol OR2C1 KP290436
1408 1274 −7 1-octanethiol OR2C1 G16S/C149W/R229H KP290437
1409 1274 −5 1-octanethiol OR2C1 P58S/C149W KP290438
1409 1274 −7 1-octanethiol OR2C1 P58S/C149W KP290438
1410 1379 −4 lyral OR10J5 R233W KP290439
1411 1416 −6 butyl anthranilate OR4Q3 KP290440
1411 1309 −6 eugenol OR4Q3 KP290440
1418 1115 −3 coumarin OR2B11 KP290447
1418 1192 −3 dicyclohexyl disulfide OR2B11 KP290447
1418 1342 −5 spearmint OR2B11 KP290447
1418 1202 −7 coffee difuran OR2B11 KP290447
1418 1360 −5 quinoline OR2B11 KP290447
1418 1111 −6 cinnamaldehyde OR2B11 KP290447
1423 1325 −4 isovaleric acid OR11H4 KP290452
1432 1326 −4 jasmine OR2T6 N21D/C23G/S243A KP290460
1457 1325 −4 isovaleric acid OR11H6 KP290483
1461 1111 −5 cinnamaldehyde OR10H2 L40Q KP290486
1461 1111 −4 cinnamaldehyde OR10H2 L40Q KP290486
1462 1324 −4 isoeugenol OR2S2 KP290487
1462 1324 −4 isoeugenol OR2S2 KP290487
1463 1299 −5 cinnamon OR1N1 KP290488
1474 1295 −3 butyric acid OR2T1 P132L KP290499
1474 1341 −5 sandalwood OR2T1 P132L KP290499
1494 1316 −3 geranyl acetate OR2M2 R220G/C235R KP290518
1494 1316 −3 geranyl acetate OR2M2 R220G/C235R KP290518
1499 1325 −3 isovaleric acid OR8U8 KP290523
1502 1278 −6 TMT OR5K1 KP290526
1502 1278 −9 TMT OR5K1 KP290526
1502 1278 −6 TMT OR5K1 KP290526
1502 1278 −9 TMT OR5K1 KP290526
1502 1278 −9 TMT OR5K1 KP290526
1502 1311 −8 eugenol methyl ether OR5K1 KP290526
1502 1278 −9 TMT OR5K1 KP290526
1505 1328 −4 linalool OR1N2 KP290529
1522 1324 −5 isoeugenol OR2AT4 KP290546
1522 1324 −5 isoeugenol OR2AT4 KP290546
1528 1024 −7 (+)-carvone OR1A1 R128H KP290552
1529 1024 −7 (+)-carvone OR1A1 KP290553
1529 1024 −8 (+)-carvone OR1A1 KP290553
1530 1024 −7 (+)-carvone OR1A1 V233M KP290554
1530 1024 −8 (+)-carvone OR1A1 V233M KP290554
1531 1024 −7 (+)-carvone OR1A1 P285S KP290555
1531 1024 −8 (+)-carvone OR1A1 P285S KP290555
1532 1325 −4 isovaleric acid OR51E1 S11N KP290556
1532 1325 −5 isovaleric acid OR51E1 S11N KP290556
1532 1325 −5 isovaleric acid OR51E1 S11N KP290556
1532 1325 −4 isovaleric acid OR51E1 S11N KP290556
1533 1191 −8 allyl phenylacetate OR51L1 T196I/A207V KP290557
1534 1191 −8 allyl phenylacetate OR51L1 KP290558
1535 1191 −7 allyl phenylacetate OR51L1 I281M KP290559
1536 1300 −6 cis-3-hexen-1-ol OR2W1 D296N KP290560
1571 1111 −5 cinnamaldehyde OR2C1 G16S/C149W/C169Y/R229H KP290588
1571 1337 −4 phenyl acetaldehyde OR2C1 G16S/C149W/C169Y/R229H KP290588
1571 1274 −4 1-octanethiol OR2C1 G16S/C149W/C169Y/R229H KP290588
1572 1300 −5 cis-3-hexen-1-ol OR2W1 M81V KP290589
1573 1325 −5 isovaleric acid OR51E1 K299R KP290590
1573 1325 −5 isovaleric acid OR51E1 K299R KP290590
1574 1290 −5 androstenone OR7D4 KP290591
1575 1300 −4 cis-3-hexen-1-ol OR2J1 L12I KP290592
1581 1388 −5 3-phenyl propyl propionate OR10A6 V140G/L287P KP290597
1581 1290 −5 androstenone OR10A6 V140G/L287P KP290597
1581 1391 −4 amyl laurate OR10A6 V140G/L287P KP290597
1581 1295 −5 butyric acid OR10A6 V140G/L287P KP290597
1581 1315 −4 androstadienone OR10A6 V140G/L287P KP290597
1581 1388 −8 3-phenyl propyl propionate OR10A6 V140G/L287P KP290597
1581 1290 −7 androstenone OR10A6 V140G/L287P KP290597
1581 1324 −4 isoeugenol OR10A6 V140G/L287P KP290597
1582 1290 −3 androstenone OR10A6 A117V/V140G/L287P KP290598
1582 1290 −4 androstenone OR10A6 A117V/V140G/L287P KP290598
1585 1310 −4 eugenol acetate OR10G3 KP290601
1585 1346 −4 vanillin OR10G3 KP290601
1589 1292 −5 banana OR10G7 KP290605
1589 1326 −6 jasmine OR10G7 KP290605
1589 1342 −4 spearmint OR10G7 KP290605
1589 1309 −6 eugenol OR10G7 KP290605
1589 1311 −6 eugenol methyl ether OR10G7 KP290605
1589 1287 −6 2-methoxy-4-methylphenol OR10G7 KP290605
1589 1332 −6 nutmeg OR10G7 KP290605
1589 1309 −6 eugenol OR10G7 KP290605
1589 1317 −6 guaiacol OR10G7 KP290605
1590 1309 −6 eugenol OR10G7 T90A KP290606
1590 1309 −6 eugenol OR10G7 T90A KP290606
1591 1309 −6 eugenol OR10G7 T13M KP290607
1591 1309 −6 eugenol OR10G7 T13M KP290607
1591 1309 −8 eugenol OR10G7 T13M KP290607
1593 1111 −6 cinnamaldehyde OR10H2 KP290609
1593 1316 −3 geranyl acetate OR10H2 KP290609
1593 1111 −4 cinnamaldehyde OR10H2 KP290609
1596 1309 −5 eugenol OR10H5 KP290612
1597 1309 −4 eugenol OR10H5 KP290613
1603 1299 −5 cinnamon OR1N1 P18S KP290618
1604 1316 −8 geranyl acetate OR2A25 KP290619
1605 1316 −5 geranyl acetate OR2A25 S75N KP290620
1606 1316 −8 geranyl acetate OR2A25 A209P KP290621
1607 1360 −6 quinoline OR2A25 S75N/A209P KP290622
1607 1316 −8 geranyl acetate OR2A25 S75N/A209P KP290622
1607 1316 −7 geranyl acetate OR2A25 S75N/A209P KP290622
1609 1202 −4 coffee difuran OR2B11 V198M KP290624
1609 1360 −5 quinoline OR2B11 V198M KP290624
1609 1342 −5 spearmint OR2B11 V198M KP290624
1609 1115 −3 coumarin OR2B11 V198M KP290624
1611 1360 −5 quinoline OR2B11 V198M/T293I/D300G KP290626
1611 1115 −4 coumarin OR2B11 V198M/T293I/D300G KP290626
1611 1342 −6 spearmint OR2B11 V198M/T293I/D300G KP290626
1611 1202 −4 coffee difuran OR2B11 V198M/T293I/D300G KP290626
1612 1111 −4 cinnamaldehyde OR2B11 I130S/V198M KP290627
1616 1300 −5 cis-3-hexen-1-ol OR2J3 I228V/M261I KP290631
1616 1111 −6 cinnamaldehyde OR2J3 I228V/M261I KP290631
1616 1334 −5 octyl aldehyde OR2J3 I228V/M261I KP290631
1616 1300 −5 cis-3-hexen-1-ol OR2J3 I228V/M261I KP290631
1616 1111 −5 cinnamaldehyde OR2J3 I228V/M261I KP290631
1616 1316 −5 geranyl acetate OR2J3 I228V/M261I KP290631
1616 1342 −5 spearmint OR2J3 I228V/M261I KP290631
1616 1311 −5 eugenol methyl ether OR2J3 I228V/M261I KP290631
1616 1300 −6 cis-3-hexen-1-ol OR2J3 I228V/M261I KP290631
1617 1300 −4 cis-3-hexen-1-ol OR2J3 I228V KP290632
1617 1300 −5 cis-3-hexen-1-ol OR2J3 I228V KP290632
1617 1308 −4 ethylene brassylate OR2J3 I228V KP290632
1617 1111 −4 cinnamaldehyde OR2J3 I228V KP290632
1617 1111 −6 cinnamaldehyde OR2J3 I228V KP290632
1617 1316 −5 geranyl acetate OR2J3 I228V KP290632
1618 1316 −3 geranyl acetate OR2J3 KP290633
1619 1316 −4 geranyl acetate OR2J3 R226Q/I228V/M261I KP290634
1619 1300 −4 cis-3-hexen-1-ol OR2J3 R226Q/I228V/M261I KP290634
1619 1111 −6 cinnamaldehyde OR2J3 R226Q/I228V/M261I KP290634
1619 1300 −4 cis-3-hexen-1-ol OR2J3 R226Q/I228V/M261I KP290634
1619 1111 −4 cinnamaldehyde OR2J3 R226Q/I228V/M261I KP290634
1623 1295 −4 butyric acid OR2T1 KP290638
1624 1295 −4 butyric acid OR2T1 I76V KP290639
1634 1299 −5 cinnamon OR3A1 KP290642
1635 1307 −4 ethyl vanillin OR3A1 R125Q KP290643
1635 1299 −5 cinnamon OR3A1 R125Q KP290643
1636 1299 −5 cinnamon OR3A1 KP290644
1638 1310 −4 eugenol acetate OR4D2 L29I KP290646
1638 1310 −4 eugenol acetate OR4D2 L29I KP290646
1639 1310 −4 eugenol acetate OR4D2 KP290647
1639 1310 −4 eugenol acetate OR4D2 KP290647
1650 1309 −6 eugenol OR4Q3 F238L KP290654
1661 1295 −4 butyric acid OR5AL1 Insertion KP290661
1666 1295 −4 butyric acid OR5D14 Q102L/S249A KP290665
1670 1311 −3 eugenol methyl ether OR6F1 F215L KP290668
1673 1295 −4 butyric acid OR7G2 V263A/F281V KP290671
1679 1310 −3 eugenol acetate OR9G1 KP290677
1683 1300 −3 cis-3-hexen-1-ol OR14J1 KP290679
1686 1316 −4 geranyl acetate OR1D2 KP290682
1692 1309 −5 eugenol OR1D5 KP290686
1724 1345 −7 undecanal OR56A4 KP290707
1727 1024 −8 (+)-carvone OR8B3 H20R/Q24R/V34I/M114I KP290709
1727 1339 −6 r-carvone OR8B3 H20R/Q24R/V34I/M114I KP290709
1727 1025 −8 (−)-carvone OR8B3 H20R/Q24R/V34I/M114I KP290709
1738 1346 −5 vanillin OR10G4 KP290714
1738 1346 −5 vanillin OR10G4 KP290714
1739 1346 −4 vanillin OR10G4 KP290715
1740 1346 −5 vanillin OR10G4 K295Q KP290716
1740 1346 −5 vanillin OR10G4 K295Q KP290716
1744 1111 −3 cinnamaldehyde OR2J3 T113A/R226Q/I228V/M261I KP290717
1744 1111 −6 cinnamaldehyde OR2J3 T113A/R226Q/I228V/M261I KP290717
1745 1300 −5 cis-3-hexen-1-ol OR2J3 KP290718
1747 1078 −6 n-amyl acetate OR2J3 KP290720
1747 1111 −4 cinnamaldehyde OR2J3 KP290720
1747 1111 −6 cinnamaldehyde OR2J3 KP290720
1747 1300 −5 cis-3-hexen-1-ol OR2J3 KP290720
1761 1290 −5 androstenone OR7D4 R88W/T133M KP290723
1761 1290 −4 androstenone OR7D4 R88W/T133M KP290723
1761 1290 −4 androstenone OR7D4 R88W/T133M KP290723
1761 1290 −3 androstenone OR7D4 R88W/T133M KP290723
1762 1290 −3 androstenone OR7D4 P79L KP290724
1764 1416 −4 butyl anthranilate OR2J2 KP290725
1764 1290 −5 androstenone OR2J2 KP290725
1764 1307 −3 ethyl vanillin OR2J2 KP290725
1765 1315 −5 androstadienone OR7C1 V126I/E171K/S210P KP290726
1768 1315 −5 androstadienone OR7C1 S99G/V126I/E171K/S210P KP290729
1781 1290 −4 androstenone OR10A6 L287P KP290731
1781 1388 −8 3-phenyl propyl propionate OR10A6 L287P KP290731
1784 1300 −5 cis-3-hexen-1-ol OR2J3 KP290732
Open in a new tab

Concentration. The molarity applied to the well. Note that the no-odor condition was coded as −12 in this column.

NormLuc. The Luc/RL ratio from each well.

OR. A unique ID for each olfactory receptor clone.

Odor. A unique ID for the odorant applied to the well.

Date. The date the experiment was run in MM/DD/YY format.

Data record 4—receptors

The receptor information is presented in a tab-separated values file (Data Citation 1). Each row represents a single olfactory receptor.

OR. A unique ID for the olfactory receptor, used in Data Records 1–3.

Gene. The gene name of the olfactory receptor encoded in a given plasmid, followed by the amino acid changes from the hg19 reference sequence for the gene. For example, ‘OR6Y1 V252I’ encodes the gene OR6Y1, but while the hg19 reference sequence has a ‘V’ as the 252nd amino acid, this clone encodes an ‘I’ at position 252. Note that some plasmids were cloned from older builds of the genome, which may start at a different methionine than the current model. These differences from h19 reference may not appear here. Please consult the nucleotide sequence for a more thorough description of differences from the reference sequence.

NucleotideSeq. The nucleotide sequence for the olfactory receptor encoded in a given plasmid. Note that the rhodopsin tag is not included in this field.

Data record 5—odors

The odor information is presented in a tab-separated values file (Data Citation 1). Each row represents a single odor. Further synonyms can be found in a file which correlates all of the CIDs in PubChem with submitted synonyms (ftp://ftp.ncbi.nlm.nih.gov/pubchem/Compound/Extras/CID-Synonym-filtered.gz).

Odor. A unique ID for the odorant, used in Data Records 1–3.

CASRegistryNum. The Chemical Abstracts Service number for the odorant, when available.

OdorName. Common name for the odorant applied to the well.

CID. PubChem Compound Identification number, a non-zero integer PubChem accession identifier for a unique chemical structure, when available.

SMILES. Simplified Molecular-Input Line-Entry System string, an ASCII string identifier for a unique chemical structure.

Technical Validation

The screen included two types of negative controls. Cells transfected with each receptor clone were challenged with a no-odor stimulation (CD293 alone) to control for baseline receptor activity. Cells transfected with an empty vector were challenged with all of the tested odorants to control for nonspecific activation. All plates in the primary screen included five broadly-tuned odorant receptors and six wells transfected with Olfr544 (also known as MOR42-3 or S6) which served as a standard. Of the six wells transfected with Olfr544, three were challenged with the diluent (CD293), and three were challenged with 10 μM of nonanedioic acid. Rankings from the primary screen consistently predicted results from later screens (Figure 5). The ultimate validation of this assay is prediction of behaviour, and previous results from similar in vitro assays have been shown to predict human olfactory perception12,15,35,36.

Figure 5. Validation of the screen.

Figure 5

Open in a new tab

An ROC curve indicates that (a) the primary screen predicts odor/receptor pairs that pass the secondary screen, (b) the primary screen predicts odor/receptor pairs that pass the dose response filter, and (c) the secondary screen predicts odor/receptor pairs that pass the dose response filter.

Usage Notes

We have included R37 scripts to facilitate analysis of the data. The included R scripts, Supplementary Files 1 and 2, have been implemented as a hosted Shiny38 application (http://www.monell.org/supplemental_files/jmainland/jm0714) to facilitate browsing the data39–41. The R markdown file, Supplementary File 3, includes code to carry out routine normalization of the primary screen, fit an ANOVA to data from the secondary screen, fit a sigmoid to the dose-response data, and create Figures 2, 3 and 5 42,43.

Additional information

Table 1 is only available in the online version of this paper.

How to cite this article: Mainland, J. D. et al. Human olfactory receptor responses to odorants. Sci. Data 2:150002 doi: 10.1038/sdata.2015.2 (2015).

Supplementary Material

sdata20152-isa1.zip (356.9KB, zip)
Supplementary Table 1
sdata20152-s2.xls (11.9MB, xls)
Supplementary File 1
sdata20152-s3.txt (1.5KB, txt)
Supplementary File 2
sdata20152-s4.txt (3.6KB, txt)
Supplementary File 3
sdata20152-s5.txt (22.9KB, txt)

Acknowledgments

This work was supported by R01 DC005782, R01 DC012095, R03 DC011373, R01 DC013339, the Defense Advanced Research Project Agency RealNose Project, and an NRSA postdoctoral fellowship F32 DC008932 to J.D.M.

Footnotes

The authors declare no competing financial interest.

Data Citations

  1. Mainland J. D., Yun R. L., Zhou T., Liu W. L. L., Matsunami H. 2014. Figshare. http://dx.doi.org/10.6084/m9.figshare.979135

References

  1. Araneda R. C., Kini A. D. & Firestein S. The molecular receptive range of an odorant receptor. Nat. Neurosci. 3, 1248–1255 (2000). [DOI] [PubMed] [Google Scholar]
  2. Katada S., Hirokawa T., Oka Y., Suwa M. & Touhara K. Structural basis for a broad but selective ligand spectrum of a mouse olfactory receptor: mapping the odorant-binding site. J. Neurosci. 25, 1806–1815 (2005). [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Malnic B., Hirono J., Sato T. & Buck L. B. Combinatorial receptor codes for odors. Cell 96, 713–723 (1999). [DOI] [PubMed] [Google Scholar]
  4. Saito H., Kubota M., Roberts R., Chi Q. & Matsunami H. RTP family members induce functional expression of mammalian odorant receptors. Cell 119, 679–691 (2004). [DOI] [PubMed] [Google Scholar]
  5. Wetzel C. H. et al. Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes. J. Neurosci. 19, 7426–7433 (1999). [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Spehr M. et al. Identification of a testicular odorant receptor mediating human sperm chemotaxis. Science 299, 2054–2058 (2003). [DOI] [PubMed] [Google Scholar]
  7. Sanz G., Schlegel C., Pernollet J.-C. & Briand L. Comparison of odorant specificity of two human olfactory receptors from different phylogenetic classes and evidence for antagonism. Chem. Senses 30, 69–80 (2005). [DOI] [PubMed] [Google Scholar]
  8. Matarazzo V. et al. Functional characterization of two human olfactory receptors expressed in the baculovirus Sf9 insect cell system. Chem. Senses 30, 195–207 (2005). [DOI] [PubMed] [Google Scholar]
  9. Jacquier V., Pick H. & Vogel H. Characterization of an extended receptive ligand repertoire of the human olfactory receptor OR17-40 comprising structurally related compounds. J. Neurochem. 97, 537–544 (2006). [DOI] [PubMed] [Google Scholar]
  10. Neuhaus E. M., Mashukova A., Zhang W., Barbour J. & Hatt H. A specific heat shock protein enhances the expression of mammalian olfactory receptor proteins. Chem. Senses 31, 445–452 (2006). [DOI] [PubMed] [Google Scholar]
  11. Schmiedeberg K. et al. Structural determinants of odorant recognition by the human olfactory receptors OR1A1 and OR1A2. J. Struct. Biol. 159, 400–412 (2007). [DOI] [PubMed] [Google Scholar]
  12. Keller A., Zhuang H., Chi Q., Vosshall L. B. & Matsunami H. Genetic variation in a human odorant receptor alters odour perception. Nature 449, 468–472 (2007). [DOI] [PubMed] [Google Scholar]
  13. Menashe I. et al. Genetic elucidation of human hyperosmia to isovaleric acid. PLoS Biol. 5, e284 (2007). [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Saito H., Chi Q., Zhuang H., Matsunami H. & Mainland J. D. Odor coding by a Mammalian receptor repertoire. Sci. Signal. 2, ra9 (2009). [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mainland J. D. et al. The missense of smell: functional variability in the human odorant receptor repertoire. Nat. Neurosci. 17, 114–120 (2014). [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Adipietro K. A., Mainland J. D. & Matsunami H. Functional evolution of mammalian odorant receptors. PLoS Genet. 8, e1002821 (2012). [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Braun T., Voland P., Kunz L., Prinz C. & Gratzl M. Enterochromaffin Cells of the Human Gut: Sensors for Spices and Odorants. Gastroenterology 132, 1890–1901 (2007). [DOI] [PubMed] [Google Scholar]
  18. Fujita Y. et al. Deorphanization of Dresden G protein-coupled receptor for an odorant receptor. J. Recept. Signal Transduct. Res. 27, 323–334 (2007). [DOI] [PubMed] [Google Scholar]
  19. Mashukova A., Spehr M., Hatt H. & Neuhaus E. M. Beta-arrestin2-mediated internalization of mammalian odorant receptors. J. Neurosci. 26, 9902–9912 (2006). [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jaeger S. R. et al. A mendelian trait for olfactory sensitivity affects odor experience and food selection. Curr. Biol. 23, 1601–1605 (2013). [DOI] [PubMed] [Google Scholar]
  21. Busse D. et al. A Synthetic Sandalwood Odorant Induces Wound Healing Processes in Human Keratinocytes via the Olfactory Receptor OR2AT4. J. Invest. Dermatol. 134, 2823–2832 (2014). [DOI] [PubMed] [Google Scholar]
  22. Hallem E. A & Carlson J. R. Coding of odors by a receptor repertoire. Cell 125, 143–160 (2006). [DOI] [PubMed] [Google Scholar]
  23. Haddad R. et al. A metric for odorant comparison. Nat. Methods 5, 425–429 (2008). [DOI] [PubMed] [Google Scholar]
  24. Olsen S. R. & Wilson R. I. Lateral presynaptic inhibition mediates gain control in an olfactory circuit. Nature 452, 956–960 (2008). [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Bhandawat V., Olsen S. R., Gouwens N. W., Schlief M. L. & Wilson R. I. Sensory processing in the Drosophila antennal lobe increases reliability and separability of ensemble odor representations. Nat. Neurosci. 10, 1474–1482 (2007). [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jiang P. et al. Major taste loss in carnivorous mammals. Proc. Natl Acad. Sci. USA 109, 4956–4961 (2012). [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. The 1000 Genomes Project Consortium et al. A map of human genome variation from population-scale sequencing. Nature 467, 1061–1073 (2010). [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Genome 10K Consortium. Genome 10K: a proposal to obtain whole-genome sequence for 10,000 vertebrate species. J. Hered. 100, 659–674 (2009). [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Klabunde T. & Hessler G. Drug design strategies for targeting G-protein-coupled receptors. ChemBioChem 3, 928–944 (2002). [DOI] [PubMed] [Google Scholar]
  30. Heckman K. L. & Pease L. R. Gene splicing and mutagenesis by PCR-driven overlap extension. Nat. Protoc. 2, 924–932 (2007). [DOI] [PubMed] [Google Scholar]
  31. Zhuang H. & Matsunami H. Evaluating cell-surface expression and measuring activation of mammalian odorant receptors in heterologous cells. Nat. Protoc. 3, 1402–1413 (2008). [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zhuang H. & Matsunami H. Synergism of accessory factors in functional expression of mammalian odorant receptors. J. Biol. Chem. 282, 15284–15293 (2007). [DOI] [PubMed] [Google Scholar]
  33. Li Y. R. & Matsunami H. Activation state of the M3 muscarinic acetylcholine receptor modulates mammalian odorant receptor signaling. Sci . Signal. 4, ra1 (2011). [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Keller A., Hempstead M., Gomez I. A., Gilbert A. N. & Vosshall L. B. An olfactory demography of a diverse metropolitan population. BMC Neurosci. 13, 122 (2012). [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McRae J. F. et al. Genetic variation in the odorant receptor OR2J3 is associated with the ability to detect the “grassy” smelling odor, cis-3-hexen-1-ol. Chem. Senses 37, 585–593 (2012). [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. McRae J. F. et al. Identification of regions associated with variation in sensitivity to food-related odors in the human genome. Curr. Biol. 23, 1596–1600 (2013). [DOI] [PubMed] [Google Scholar]
  37. R Development Core Team. R: A Language and Environment for Statistical Computing http://www.r-project.org (2009).
  38. RStudio and Inc. Shiny: Web Application Framework for R http://cran.r-project.org/package=shiny (2014).
  39. Wickham H. ggplot2: elegant graphics for data analysis. Media 35 (Springer, 2009). [Google Scholar]
  40. Wickham H. The Split-Apply-Combine strategy for data. J. Stat. Softw. 40, 1–29 (2011). [Google Scholar]
  41. Ritz C., Streibig J. C., Ritz C. & Streibig J. C. Bioassay Analysis using R. J. Stat. Softw. 12, 1–22 (2005). [Google Scholar]
  42. Robin X. et al. pROC: an open-source package for R and S+ to analyze and compare ROC curves. BMC Bioinformatics 12, 77 (2011). [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wickham H. Reshaping Data with the reshape Package. J. Stat. Softw. 21, 1–20 (2007). [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Citations

  1. Mainland J. D., Yun R. L., Zhou T., Liu W. L. L., Matsunami H. 2014. Figshare. http://dx.doi.org/10.6084/m9.figshare.979135

Supplementary Materials

sdata20152-isa1.zip (356.9KB, zip)
Supplementary Table 1
sdata20152-s2.xls (11.9MB, xls)
Supplementary File 1
sdata20152-s3.txt (1.5KB, txt)
Supplementary File 2
sdata20152-s4.txt (3.6KB, txt)
Supplementary File 3
sdata20152-s5.txt (22.9KB, txt)

Articles from Scientific Data are provided here courtesy of Nature Publishing Group

RESOURCES