Direct NMR Detection of the Binding of Functional Ligands to the Human Sweet Receptor, a Heterodimeric Family 3 GPCR
作者： Fariba M. Assadi-Porter , Marco Tonelli , Emeline Maillet , Klaas Hallenga , Outhiriaradjou Benard
DOI: 10.1021/JA8016939
关键词: Receptor 、 Membrane 、 Chemistry 、 Biochemistry 、 Mechanism of action 、 Nuclear magnetic resonance spectroscopy 、 Brazzein 、 Alitame 、 G protein-coupled receptor 、 Mutant
摘要: We present a robust method for monitoring the binding of ligands to heterodimeric (T1R2+T1R3) human sweet receptor (a family 3 GPCR receptor). The approach utilizes saturation transfer difference (STD) NMR spectroscopy with proteins expressed on surface epithelial kidney cells. preparation investigated by can contain either live cells or membranes isolated from these containing receptor. have used this confirm noncompetitive alitame and cyclamate determine that greatly reduced affinity compared wild-type brazzein explains lack sweetness mutant A16C17. This opens new avenues research mechanism action design noncalorigenic sweeteners.
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