Direct NMR detection of the binding of functional ligands to the human sweet receptor, a heterodimeric family 3 GPCR

被引：56

作者：

Assadi-Porter, Fariba M. ^{[1
]}

Tonelli, Marco ^{[1
]}

Maillet, Emeline ^{[2
]}

Hallenga, Klaas ^{[1
]}

Benard, Outhiriaradjou ^{[2
]}

Max, Marianna ^{[2
]}

Markley, John L. ^{[1
]}

机构：

[1] Univ Wisconsin, Dept Biochem, Natl Magnet Resonance Facil, Madison, WI 53706 USA

[2] Mt Sinai Sch Med, Dept Biophys, New York, NY 10029 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2008年 / 130卷 / 23期

关键词：

D O I：

10.1021/ja8016939

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

We present a robust method for monitoring the binding of ligands the heterodimeric (T1R2+T1R3) human sweet receptor (a family 3 GPCR receptor). The approach utilizes saturation transfer (STD) NMR spectroscopy with receptor proteins expressed on the surface of human epithelial kidney cells. The preparation investigated by NMR can contain either live cells or membranes isolated from these cells containing the receptor. We have used approach to confirm the noncompetitive binding of alitame and cyclamate to the receptor and to determine that greatly reduced receptor binding affinity compared to wild-type brazzein explains the lack of sweetness of brazzein mutant A16C17. This approach opens new avenues for research on the mechanism of action of the sweet receptor and for the design of new noncalorigenic sweeteners.

引用

页码：7212 / +

页数：3

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