Verveer P J, Wouters F S, Reynolds A R, Bastiaens P I
Cell Biology and Cell Biophysics Program, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Science. 2000 Nov 24;290(5496):1567-70. doi: 10.1126/science.290.5496.1567.
Evidence for a new signaling mechanism consisting of ligand-independent lateral propagation of receptor activation in the plasma membrane is presented. We visualized the phosphorylation of green fluorescent protein (GFP)-tagged ErbB1 (ErbB1-GFP) receptors in cells focally stimulated with epidermal growth factor (EGF) covalently attached to beads. This was achieved by quantitative imaging of protein reaction states in cells by fluorescence resonance energy transfer (FRET) with global analysis of fluorescence lifetime imaging microscopy (FLIM) data. The rapid and extensive propagation of receptor phosphorylation over the entire cell after focal stimulation demonstrates a signaling wave at the plasma membrane resulting in full activation of all receptors.
本文提出了一种新的信号传导机制的证据，该机制由质膜中受体激活的非配体依赖性横向传播组成。我们通过共价连接到珠子上的表皮生长因子（EGF）对细胞进行局部刺激，观察了绿色荧光蛋白（GFP）标记的ErbB1（ErbB1-GFP）受体的磷酸化情况。这是通过荧光共振能量转移（FRET）对细胞中的蛋白质反应状态进行定量成像，并对荧光寿命成像显微镜（FLIM）数据进行全局分析来实现的。局部刺激后，受体磷酸化在整个细胞中迅速而广泛地传播，这表明质膜上存在信号波，导致所有受体完全激活。
