Live‐cell imaging of endogenous Ras‐GTP illustrates predominant Ras activation at the plasma membrane
作者： Martin Augsten , Rico Pusch , Christoph Biskup , Knut Rennert , Ute Wittig
关键词: GTP' 、 Golgi apparatus 、 Effector 、 Biology 、 Green fluorescent protein 、 Endogeny 、 Jurkat cells 、 Molecular biology 、 Cell biology 、 Anti-apoptotic Ras signalling cascade 、 Live cell imaging
摘要: Ras-GTP imaging studies using the Ras-binding domain (RBD) of Ras effector c-Raf as a reporter for overexpressed have produced discrepant results about possible activation at Golgi apparatus. We report that RBD oligomerization provides probes visualization endogenous Ras-GTP, obviating overexpression and side effects derived thereof. in tenacious binding to interruption signalling. Trimeric fused green fluorescent protein agonist-induced plasma membrane (PM) COS-7, PC12 Jurkat cells, but do not accumulate Golgi. PM illumination is exacerbated by its sensitivity dominant-negative RasS17N pharmacological manipulations matches formation assessed biochemically. Our data illustrate Golgi-located under control growth factors argue predominant site activation.
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