Suppression of hTAS2R16 Signaling by Umami Substances
Abstract
:1. Introduction
2. Results and Discussion
2.1. Effect of MSG, IMP, and l-Theanine on Ligand Binding to the hTAS2R16
2.2. The Inhibitory Potency of Umami Substances on Salicin Binding to the hTAS2R16
2.3. Mechanism of Antagonism of the hTAS2R16 by Umami Substances
2.4. Synergism between Umami Substances on the Suppression of Bitterness
2.5. Point Mutant Analysis Using Two Known Probenecid-Insensitive Mutants, hTAS2R16 N96T and P44T, for the Potential Binding Sites of Umami Substances
3. Materials and Methods
3.1. Materials
3.2. Cell Culture and Transfection
3.3. Measurement of Cellular Ca2+ Responses in HEK293 Cells Expressing Wild-Type and Mutant hTAS2R16 by Cell-Based Assay
3.4. Measurement of Cellular Ca2+ Responses in HEK293T Cells Expressing hTAS2R by Calcium Imaging Analysis
3.5. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Salicin (0–30 mM) + Sample (mM) | EC50 (mM) | Maximal Response (ΔF/F0) | |
---|---|---|---|
+ Probenecid | 0 | 0.90 ± 0.06 † | 0.54 |
0.1 | 0.98 ± 0.10 | 0.47 | |
0.3 | 1.07 ± 0.11 | 0.23 | |
1 | NE ‡ | 0.07 | |
+ Glu-Glu | 0 | 1.04 ± 0.03 | 0.54 |
1 | 1.18 ± 0.05 | 0.51 | |
3 | 1.53 ± 0.06 | 0.49 | |
6 | 1.40 ± 0.11 | 0.40 | |
10 | 1.96 ± 0.15 | 0.33 | |
+ MSG | 0 | 1.04 ± 0.02 | 0.51 |
10 | 1.12 ± 0.05 | 0.55 | |
30 | 1.06 ± 0.03 | 0.54 | |
80 | 2.13 ± 0.05 | 0.49 | |
100 | 8.83 ± 0.09 | 0.47 | |
+ IMP | 0 | 0.92 ± 0.03 | 0.54 |
1 | 0.97 ± 0.04 | 0.53 | |
10 | 1.73 ± 0.06 | 0.53 | |
20 | 1.45 ± 0.13 | 0.40 | |
30 | 1.95 ± 0.09 | 0.27 | |
+ l-Theanine | 0 | 0.89 ± 0.05 | 0.56 |
10 | 0.78 ± 0.06 | 0.53 | |
30 | 1.00 ± 0.04 | 0.50 | |
80 | 1.65 ± 0.14 | 0.43 | |
100 | 2.74 ± 0.17 | 0.31 |
10 mM Salicin + Sample | + IMP (mM) | IC50 (mM) |
---|---|---|
+ Glu-Glu | 0 | 7.25 ± 0.02 |
1 | 7.03 ± 0.01 | |
3 | 6.97 ± 0.02 | |
10 | 7.28 ± 0.02 | |
+ MSG | 0 | 64.67 ± 0.04 |
1 | 59.85 ± 0.05 | |
5 | 49.65 ± 0.06 | |
10 | 19.51 ± 0.09 | |
+ l-Theanine | 0 | 77.33 ± 0.05 |
1 | 78.13 ± 0.06 | |
3 | 79.86 ± 0.06 | |
10 | 69.62 ± 0.06 |
Sample (mM) | EC50 (mM) | Maximal Response (ΔF/F0) | |||||
---|---|---|---|---|---|---|---|
WT | N96T | P44T | WT | N96T | P44T | ||
+ Probenecid | 0 | 0.90 ± 0.04 † | 0.23 ± 0.03 | 0.64 ± 0.06 | 0.54 | 0.68 | 0.56 |
0.3 | 1.04 ± 0.19 | 0.26 ± 0.03 | 0.94 ± 0.10 | 0.23 | 0.68 | 0.58 | |
+ Glu-Glu | 0 | 1.04 ± 0.03 | 0.25 ± 0.05 | 0.62 ± 0.04 | 0.54 | 0.61 | 0.62 |
6 | 1.35 ± 0.12 | 0.24 ± 0.07 | 0.59 ± 0.07 | 0.40 | 0.65 | 0.42 | |
+ MSG | 0 | 1.07 ± 0.02 | 0.18 ± 0.04 | 0.68 ± 0.03 | 0.51 | 0.66 | 0.56 |
80 | 2.03 ± 0.06 | 0.29 ± 0.10 | 0.94 ± 0.14 | 0.49 | 0.65 | 0.57 | |
+ IMP | 0 | 0.96 ± 0.03 | 0.20 ± 0.03 | 0.69 ± 0.04 | 0.54 | 0.66 | 0.61 |
20 | 0.97 ± 0.12 | 0.30 ± 0.15 | 0.94 ± 0.13 | 0.40 | 0.28 | 0.37 | |
+ l-Theanine | 0 | 1.10 ± 0.03 | 0.20 ± 0.04 | 0.75 ± 0.04 | 0.56 | 0.63 | 0.59 |
80 | 1.73 ± 0.16 | 0.31 ± 0.07 | 1.03 ± 0.03 | 0.43 | 0.66 | 0.56 |
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Rhyu, M.-R.; Kim, Y.; Misaka, T. Suppression of hTAS2R16 Signaling by Umami Substances. Int. J. Mol. Sci. 2020, 21, 7045. https://doi.org/10.3390/ijms21197045
Rhyu M-R, Kim Y, Misaka T. Suppression of hTAS2R16 Signaling by Umami Substances. International Journal of Molecular Sciences. 2020; 21(19):7045. https://doi.org/10.3390/ijms21197045
Chicago/Turabian StyleRhyu, Mee-Ra, Yiseul Kim, and Takumi Misaka. 2020. "Suppression of hTAS2R16 Signaling by Umami Substances" International Journal of Molecular Sciences 21, no. 19: 7045. https://doi.org/10.3390/ijms21197045