Design and construction of conformational biosensors to monitor ion channel activation: A prototype FlAsH/BRET-approach to Kir3 channels
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- 作者：Derek N. Robertsona ; 1 ; Rory Slenob ; 1 ; Karim Nagia ; d ; Darlaine Pé ; trinb ; Terence E. Hé ; bertb ; Graciela Pineyroa ; c ; d ; e ; graciela.pineyro.filpo@umontreal.ca" class="auth_mail" title="E-mail the corresponding author
- 关键词：BRET ; Bioluminescence Resonance Energy Transfer ; FlAsH-EDT2 ; fluorescein arsenical hairpin binder-ethanedithiol ; TC ; tetracysteine ; Kir3/GIRK ; G protein-gated inwardly rectifying potassium channels ; RLuc ; Renilla luciferase ; NLuc ; Nano-luciferase ; δOR ; δ-opioid receptor ; CTD ; C-terminal domain ; NTD ; N-terminal domain ; TMD ; trans-membrane domain
- 刊名：Methods
- 出版年：2016
- 出版时间：1 January 2016
- 年：2016
- 卷：92
- 期：Complete
- 页码：19-35
- 全文大小：1684 K
文摘
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We design a new set of BRET biosensors based on the fluorescent properties of TC-domain/FlAsH-EDT2.
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We use the TC domain to allow us to use BRET to measure conformational changes within the channel.
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These changes allow us to measure dose response curves for channel agonists and δ-opioid receptor agonists.
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The BRET responses we see can be compared to other measures of channel activation.