ID | 59909 |
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Author |
Yoshida, Takashi
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Yasui, Norihisa
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
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Kusakabe, Yuko
Food Research Institute, National Agriculture and Food Research Organization
Ito, Chiaki
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Akamatsu, Miki
Graduate School of Agriculture, Kyoto University
Yamashita, Atsuko
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
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Abstract | Taste receptor type 1 (T1r) is responsible for the perception of essential nutrients, such as sugars and amino acids, and evoking sweet and umami (savory) taste sensations. T1r receptors recognize many of the taste substances at their extracellular ligand-binding domains (LBDs). In order to detect a wide array of taste substances in the environment, T1r receptors often possess broad ligand specificities. However, the entire ranges of chemical spaces and their binding characteristics to any T1rLBDs have not been extensively analyzed. In this study, we exploited the differential scanning fluorimetry (DSF) to medaka T1r2a/T1r3LBD, a current sole T1rLBD heterodimer amenable for recombinant preparation, and analyzed their thermal stabilization by adding various amino acids. The assay showed that the agonist amino acids induced thermal stabilization and shifted the melting temperatures (T-m) of the protein. An agreement between the DSF results and the previous biophysical assay was observed, suggesting that DSF can detect ligand binding at the orthostericbinding site in T1r2a/T1r3LBD. The assay further demonstrated that most of the tested Lamino acids, but no D-amino acid, induced T-m shifts of T1r2a/T1r3LBD, indicating the broad L-amino acid specificities of the proteins probably with several different manners of recognition. The T-m shifts by each amino acid also showed a fair correlation with the responses exhibited by the full-length receptor, verifying the broad amino-acid binding profiles at the orthosteric site in LBD observed by DSF.
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Published Date | 2019-10-04
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Publication Title |
PLOS ONE
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Volume | volume14
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Issue | issue10
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Publisher | Public Library of Science
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Start Page | e0218909
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ISSN | 1932-6203
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Content Type |
Journal Article
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language |
English
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OAI-PMH Set |
岡山大学
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Copyright Holders | © 2019 Yoshida et al.
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File Version | publisher
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DOI | |
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Related Url | isVersionOf https://doi.org/10.1371/journal.pone.0218909
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License | https://creativecommons.org/licenses/by/4.0/
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Funder Name |
Japan Society for the Promotion of Science
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助成番号 | 15H05370
17H03644
18H04621
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