ID 59909
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Yoshida, Takashi Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Yasui, Norihisa Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University ORCID Kaken ID publons researchmap
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 Kaken ID publons
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.
Published Date
2019-10-04
Publication Title
PLOS ONE
Volume
volume14
Issue
issue10
Publisher
Public Library of Science
Start Page
e0218909
ISSN
1932-6203
Content Type
Journal Article
language
英語
OAI-PMH Set
岡山大学
Copyright Holders
© 2019 Yoshida et al.
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Related Url
isVersionOf https://doi.org/10.1371/journal.pone.0218909
License
https://creativecommons.org/licenses/by/4.0/
Funder Name
Japan Society for the Promotion of Science
助成番号
15H05370
17H03644
18H04621