FRET analysis reveals a critical conformational change within the Na,K-ATPase alpha1 subunit N-terminus during GPCR-dependent endocytosis |
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Authors: | Efendiev Riad Cinelli Angel R Leibiger Ingo B Bertorello Alejandro M Pedemonte Carlos H |
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Affiliation: | Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, 4800 Calhoun Road, Houston, TX 77204-5037, USA. refendiev@uh.edu |
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Abstract: | Dopamine is a major regulator of sodium reabsorption in proximal tubule epithelia. It induces the endocytosis of plasma membrane Na,K-ATPase molecules, and this results in a reduced capacity of the cells to transport sodium. Dopamine induces the phosphorylation of Ser-18 in the alpha1-subunit of Na,K-ATPase. Fluorescence resonance energy transfer analysis of cells expressing YFP-alpha1 and beta1-CFP reveals that treatment of the cells with dopamine increases energy transfer between CFP and YFP. This is consistent with a protein conformational change that results in the N-terminal end of alpha1 moving closer to the internal face of the plasma membrane. |
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Keywords: | CFP, cyan fluorescent protein DA, dopamine NKA, Na,K-ATPase FRET, fluorescence resonance energy transfer OK, opossum kidney PI3K, phosphoinositide-3′ kinase PKC, protein kinase C YFP, yellow fluorescent protein |
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