Novel Green Fluorescent Protein-Based Ratiometric Indicators for Monitoring pH in Defined Intracellular Microdomains
作者： Takeo Awaji , Akira Hirasawa , Hideki Shirakawa , Gozoh Tsujimoto , Shunichi Miyazaki
关键词:
摘要: To measure pH in defined intracellular microdomains of living cells, we developed ratiometric indicators based on fusing tandem two green fluorescent protein (GFP) variants having different sensitivities. The function a single-excitation/dual-emission mode involving fluorescence resonance energy transfer, as well dual-excitation/single-emission mode. ratio from GFpH and YFpH showed dependency pK(a) values were 6.1 6.8, respectively. Using these expressed cultured measured visualized changes the cytosol nucleus. Furthermore, by tethering indicator to membrane (the alpha(1B) adrenergic receptor), vicinity during internalization caused endocytosis after agonist stimulation. These novel probes will serve useful tool for monitoring organelle microenvironment target protein, analyze cellular function.
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