2012 Fiscal Year Final Research Report
Elucidation of the structure-function relationships in sweet-tasting proteins by x-ray analysis at atomic resolution
| Project/Area Number |
22580105
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Kyoto University |
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Principal Investigator |
MASUDA Tetsuya 京都大学, 大学院・農学研究科, 助教 (80311744)
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| Project Period (FY) |
2010 – 2012
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| Keywords | 甘味タンパク質 / ソーマチン / X線結晶構造解析 / 甘味受容体 |
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| Research Abstract |
The crystal structure of a recombinant thaumatin I was determined to a resolution of 1.1 A (PDB:3AL7). The crystallographic structure of one of its variants, thaumatin II, was determined at a resolution of 1.27 A (PDB:3AOK). The results showed the critical positive charges are disordered and the flexibility and fluctuation of these side chains would be suitable for interaction with sweet receptors. The atomic structure of a recombinant thaumatin at pH 8.0 was determined at a resolution of 1.0 A (PDB:3VJQ). The results suggested a striking increase in the mobility of some lysine residues, which could facilitate a reaction with a free sulfhydryl residue produced via the β-elimination of disulfide bonds by heating at a pH above 7.0. Various mutants of thaumatin were prepared and their sweetness were quantitatively evaluated by cell-based assays using HEK293 cells expressing human sweet receptors. Chimeric human- mouse sweet receptors were constructed and their responses to sweeteners were investigated. The results showed the cysteine-rich domain of human T1R3 is important for the response to thaumatin.
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Research Products
(32 results)
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[Journal Article] Bacterial Heat Shock Protein 60, GroEL, Can Induce the Conversion of Naive T Cells into a CD4+ CD25+ Foxp3-expressing Phenotype2011
Author(s)
Ohue, R., Hashimoto, K., Nakamoto, M., Furukawa, Y., Masuda, T., Kitabatake, N., and Tani, F
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Journal Title
J. Innate Immunology
Volume: 3
Pages: 605-613
DOI
Peer Reviewed
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[Presentation] Properties of the sweet-tasting protein, thaumatin2012
Author(s)
Masuda, T., Sano, A., Ohta, K., Taguchi, W., Tsukahara, A., Tani, F., and Kitabatake, N
Organizer
World Congress on Oleo Science & 29th ISF Congress
Place of Presentation
Arkas Sasebo, Nagasaki, Japan
Year and Date
20120930-1004
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[Remarks] 甘味タンパク質の機能発現に関する高分解能X線結晶構造解析、SPring-8 利用報告書、2012B1067
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[Remarks] 高分解能X線結晶構造解析による甘味タンパク質の甘味発現機構の解明、SPring-8 利用報告書、2012A1048
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[Remarks] 甘味タンパク質の機能発現に関する高分解能構造解析、SPring-8 利用報告書、2011B1073
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[Remarks] 味覚関連タンパク質の機能発現に関する高分解能構造解析、SPring-8 利用報告書、2010B1064
