Using Bioluminescence Resonance Energy Transfer to measure ion channel assembly.
Bioluminescence Resonance Energy Transfer (BRET) measures protein interactions within 10 nm of each other. Aside from its ability to probe for interactions at high resolution, this technique operates in live, intact cells, and offers a high throughput method of detection. Thus far, BRET has been widely used in measuring G protein receptor dimerization. In this chapter, we describe the BRET methodology in detail and apply this technique to the measurement of ion channel assembly. In addition, we discuss how BRET can be used to compare the extent of homomeric and heteromeric channel assembly.
Whitaker
Gina M
GM
Department of Cellular and Physiological Sciences, University of British Columbia, Vancouver, BC, Canada.
Accili
Eric A
EA
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D019204:GTP-Binding Proteins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007473:Ion Channels / Q000096:biosynthesis / Q000502:physiology*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D014162:Transfection
D054815:Cyclic Nucleotide-Gated Cation Channels; D007473:Ion Channels; D049452:Green Fluorescent Proteins; D008156:Luciferases; D019204:GTP-Binding Proteins