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Characterization of a CFTR construct with a C-terminal tetracysteine sequence and its use in the visualization of trafficking pathways.

التفاصيل البيبلوغرافية
العنوان: Characterization of a CFTR construct with a C-terminal tetracysteine sequence and its use in the visualization of trafficking pathways.
المؤلفون: Malik, Firhan A.1, Bear, Christine E.1
المصدر: FASEB Journal. Apr2007, Vol. 21 Issue 5, pA243-A243. 1/5p.
مصطلحات موضوعية: *PROTEINS, *CYSTIC fibrosis, *ENDOCYTOSIS, *FLUORESCEIN, *GENE expression
مستخلص: Membrane permeable bi-arsenical dyes, such as fluorescein arsenical hairpin (FlAsH), bind tetracysteine-tagged-proteins emitting fluorescence in-vivo. This fluorescence method (developed by R. Tsien) provides a tool to evaluate trafficking pathways in living cells. We generated a tetracysteine-tagged Cystic Fibrosis Transmembrane Regulator (CFTR) protein and studied its expression with the long-term goal of using this protein to study its trafficking during endocytosis. The tetracysteine-tagged CFTR (CFTR-TC) was predominantly expressed in both BHK and in Cos-7 cells as the immature core-glycosylated form (band B) rather than the mature glycosylated band C. Iodide efflux assays of CFTR-TC's function as a chloride channel at the cell surface revealed an insignificant response after cAMP stimulation, likely reflecting its low band C expression. Interestingly, 1 hour FlAsH pretreatment evoked a mean response difference of -16.0 mV versus untreated (one-way ANOVA p<0.05), suggesting increased band C expression. Our results suggest that FlAsH rescues the surface expression of CFTR-TC and studies are ongoing to visualize its localization and the regulation of its trafficking by proteins involved in the endocytic pathway. [ABSTRACT FROM AUTHOR]
قاعدة البيانات: Academic Search Index
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