期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 276, 期 23, 页码 19793-19799出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M100621200
关键词
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The NK1 neurokinin receptor presents two non-ideal binding phenomena, two-component binding curves for all agonists and significant differences between agonist affinity determined by homologous versus heterologous competition binding. Whole cell binding with fusion proteins constructed between either G alpha (s) or G alpha (q), and the NK1 receptor with a truncated tail, which secured non-promiscuous G-protein interaction, demonstrated monocomponent agonist binding closely corresponding to either of the two affinity states found in the wild-type receptor. High affinity binding of both substance P and neurokinin A was observed in the tail-truncated G alpha (s) fusion construct, whereas the lower affinity component was displayed by the tail-truncated G alpha (q) fusion. The elusive difference between the affinity determined in heterologous versus homologous binding assays for substance P and especially for neurokinin A was eliminated in the G-protein fusions, An NK1 receptor mutant with a single substitution at the extracellular end of TM-III(F111S), which totally uncoupled the receptor from G alpha (s) signaling, showed binding properties that were monocomponent and otherwise very similar to those observed in the tail-truncated G alpha (q) fusion construct. Thus, the heterogenous pharmacological phenotype displayed by the NK1 receptor is a reflection of the occurrence of two active conformations or molecular phenotypes representing complexes with the G alpha (s) and G alpha (q) species, respectively. We propose that these molecular forms do not interchange readily, conceivably because of the occurrence of microdomains or signal-transductosomes within the cell membrane.
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