Use of Kikume green-red fusions to study the influence of pharmacological chaperones on trafficking of G protein-coupled receptors

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• 作者：Ingrid Ridelis^a ; ^b ; Antje Schmidt^a ; Anke Teichmann^a ; Jens Furkert^a ; Burkhard Wiesner^a ; Ralf Schü ; lein^a ; ^{schuelein@fmp-berlin.de}
• 关键词：Photoconvertible fluorescent proteins ; G protein-coupled receptor ; Vasopressin V2 receptor ; Pharmacological chaperone ; Post-translational rescue ; Co-translational rescue
• 刊名：FEBS Letters
• 出版年：2012
• 期刊代码：70_00145793
• 类别：bio
• 出版时间：23 March, 2012
• 卷：586
• 期：6
• 页码：784-791
• 文件大小：850 K

摘要

In this study we demonstrate that the photoconvertible monomeric Kikume green-red (mKikGR) protein is suitable to study trafficking of G protein-coupled receptors. Taking mKikGR-tagged mutants of the vasopressin V₂ receptor (V₂R) as models, we analyzed whether the V₂R-specific pharmacological chaperone SR121463B influences receptor folding on a co- or post-translational level. Misfolded mKikGR-tagged V₂Rs were completely photoconverted in the early secretory pathway yielding a red receptor population (already synthesized receptors) and an arising green receptor population (newly synthesized receptors). Trafficking of both receptor populations could be rescued by treatment with SR121463B demonstrating that the substance can act co- and post-translationally.