کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1938788 1050747 2006 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Co-localization of the α-subunit of BK-channels and c-PLA2 in GH3 cells
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Co-localization of the α-subunit of BK-channels and c-PLA2 in GH3 cells
چکیده انگلیسی

Large conductance, calcium-activated potassium channels (maxi K- or BK-channels) can be regulated by arachidonic acid produced by c-Phospholipase A2 (c-PLA2). Since in every excised patch from GH3 cells where there was BK-channel activity, treatment with either a stimulator or inhibitor of c-PLA2 resulted in a corresponding increase or decrease in BK-channel activity, we hypothesized that there must be a close association between BK-channel proteins and c-PLA2 in the cell membrane. To test this hypothesis, we first determined whether the two proteins would co-immunoprecipitate. We then used confocal imaging of fluorescently tagged proteins to determine where in the cells BK-channel proteins and c-PLA2 co-localize. The α-subunit of the BK-channel was strongly co-immunoprecipitated by c-PLA2 antibodies, suggesting that most of the BK channel α-subunits are associated with c-PLA2. This interaction was not affected by pharmacologically inhibiting c-PLA2 suggesting that the association does not require functionally active c-PLA2. Following dual immunohistochemical labeling and confocal microscopy, image analysis revealed that in the cytosol there was some co-localization, but most of the c-PLA2 was separate from BK-channel proteins. On the other hand, the c-PLA2 and BK-channel proteins at the plasma membrane were strongly co-localized. Immunoprecipitation experiments conducted with plasma membrane proteins support these findings. We conclude that c-PLA2 is likely physically associated with BK-channel proteins at the cell surface.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 350, Issue 1, 10 November 2006, Pages 39–49
نویسندگان
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