pone.0201210.g007.tif (1.87 MB)
The α1-EGFP/β2 receptor cannot be solubilized by detergent from Sf9 cells.
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posted on 2018-07-20, 17:36 authored by Derek P. Claxton, Eric Gouaux(A) Diagram of insect cell expression vectors harboring one (pFastBac1) or both (pFastBac Dual) full-length GABAA subunits. EGFP has been placed in the M3/M4 loop of the α1 subunit. (B) EGFP fluorescence is observed 48 hours post infection for pFastBac1 or pFastBac Dual constructs, suggesting receptor targeting to the cell membrane. (C) FSEC analysis at the indicated time points to measure expression suggests that most of the receptor breaks down upon whole cell solubilization with C12M detergent. (D) FSEC profiles do not improve using the bicistronic pFastBac Dual vector, which ensures each cell possesses a copy of both subunits. Absolute fluorescence intensities for (C) and (D) are shown on the same scale.
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GABA-gated chloride channelsshape ion channel propertiesreceptor subunit assembliesα1 subunit isoformaffinity agonist binding activityGABA-gated channel activityassemblyFSECN-linked glycosylation sitesRobust heterologous expressionreceptor expressionCys-loop receptor familyfluorescence detection size exclusion chromatographyGFP
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