دورية أكاديمية
N-terminal tagging of human P2X7 receptor disturbs calcium influx and dye uptake
| العنوان: | N-terminal tagging of human P2X7 receptor disturbs calcium influx and dye uptake |
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| المؤلفون: | Dreisig, Karin, Kristensen, Nikolaj Pagh, Dommer, Maja Wallentin, Jørgensen, Niklas Rye, Kornum, Birgitte Rahbek |
| المصدر: | Dreisig , K , Kristensen , N P , Dommer , M W , Jørgensen , N R & Kornum , B R 2018 , ' N-terminal tagging of human P2X7 receptor disturbs calcium influx and dye uptake ' , Purinergic Signalling , vol. 14 , no. 1 , pp. 83–90 . https://doi.org/10.1007/s11302-017-9598-8Test |
| سنة النشر: | 2018 |
| المجموعة: | University of Southern Denmark: Research Output / Syddansk Universitet |
| مصطلحات موضوعية: | ATP, Flow cytometry, Fluo4, Purinergic, Green Fluorescent Proteins/pharmacology, Receptors, Purinergic P2X7/metabolism, Humans, Calcium Channels/metabolism, Signal Transduction/drug effects, Fluorescent Dyes/pharmacology |
| الوصف: | The P2X7 receptor is a frequently studied member of the purinergic receptor family signalling via channel opening and membrane pore formation. Fluorescent imaging is an important molecular method for studying cellular receptor expression and localization. Fusion of receptors to fluorescent proteins might cause major functional changes and requires careful functional evaluation such as has been done for the rat P2X7 receptor. This study examines fusion constructs of the human P2X7 receptor. We assessed surface expression, channel opening with calcium influx, and pore formation using YO-PRO-1 dye uptake in response to BzATP stimulation in transfected cells. We found that tagging at the N-terminal of the human P2X7 receptor with the enhanced green fluorescent protein (eGFP) disturbed channel opening and pore formation despite intact surface expression. A triple hemagglutinin (3HA) fused to the N-terminal also disrupted pore formation but not channel opening showing that even a small tag alters the normal function of the receptor. Together, this suggests that in contrast to what has been observed for the rat P2X7 receptor, the human P2X7 receptor contains N-terminal motifs important for signalling that prevent the construction of a functionally active fusion protein. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| العلاقة: | https://portal.findresearcher.sdu.dk/da/publications/a7dc844a-09e7-44ef-94b0-c4cd4bdac8bbTest |
| DOI: | 10.1007/s11302-017-9598-8 |
| الإتاحة: | https://doi.org/10.1007/s11302-017-9598-8Test https://portal.findresearcher.sdu.dk/da/publications/a7dc844a-09e7-44ef-94b0-c4cd4bdac8bbTest https://findresearcher.sdu.dk/ws/files/151867161/11302_2017_Article_9598.pdfTest |
| حقوق: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.94C96F2E |
| قاعدة البيانات: | BASE |
| DOI: | 10.1007/s11302-017-9598-8 |
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