研究GPCR信号传导中内体cAMP产生的调控。
Studying the regulation of endosomal cAMP production in GPCR signaling.
作者信息
Gidon Alexandre, Feinstein Timothy N, Xiao Kunhong, Vilardaga Jean-Pierre
机构信息
Molecular Mechanisms of Mycobacterial Infection, Center for Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, Norway.
Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
出版信息
Methods Cell Biol. 2016;132:109-26. doi: 10.1016/bs.mcb.2015.10.007. Epub 2015 Dec 24.
Abstract
We describe methods based on live cell fluorescent microscopy and mass spectrometry to characterize the mechanism of endosomal cAMP production and its regulation using the parathyroid hormone (PTH) type 1 receptor as a prime example. These methods permit to measure rapid changes of cAMP levels in response to PTH, kinetics of endosomal ligand-receptor interaction, pH changes associated with receptor trafficking, and to identify the endosomal receptor interactome.
摘要
我们描述了基于活细胞荧光显微镜和质谱的方法，以甲状旁腺激素（PTH）1型受体为例，来表征内体cAMP产生的机制及其调控。这些方法能够测量cAMP水平对PTH的快速变化、内体配体-受体相互作用的动力学、与受体运输相关的pH变化，并鉴定内体受体相互作用组。
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