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首页> 外文期刊>Current Biology: CB >EGF regulation of PITP dynamics is blocked by inhibitors of phospholipase C and of the Ras-MAP kinase pathway
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EGF regulation of PITP dynamics is blocked by inhibitors of phospholipase C and of the Ras-MAP kinase pathway

机译：EGF对PITP动力学的调节被磷脂酶C和Ras-MAP激酶途径的抑制剂阻断

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摘要

Phosphatidylinositol transfer proteins (PITP) function in signal transduction and in membrane traffic [1, 2]. Studies aimed at elucidating the mechanism of action of PITP have yielded a singular theme; the activity of PITP stems from its ability to transfer phosphatidylinositol (PI) from its site of synthesis to sites of cellular activity and to stimulate the local synthesis of phosphorylated forms of PI. The participation of various phosphoinositides in EGF signal transduction and in the trafficking of the EGF receptors is well documented [3]. Using fluorescence lifetime imaging microscopy (FLIM) to measure fluorescence resonance energy transfer (FRET) between EGFP-PITP proteins and fluorescently labeled phospholipids, we report that PITPalpha and PITPbeta can dynamically interact with PI or PC at the plasma membrane when stimulated with EGF. Additionally, PITPbeta is localized at the Golgi, and EGF stimulation resulted in enhanced FRET. Inhibitors of the PLC and the Ras/MAP kinase pathway were both able to inhibit the EGF-stimulated interaction of PITP(x with PI at the plasma membrane. The mobility of PITP proteins was determined by using fluorescence recovery after photobleaching (FRAP), and EGF stimulation reduced the mobility at the plasma membrane. We conclude that the dynamic behavior of PITPa. and PITPbeta in vivo is a regulated process involving multiple mechanisms.
机译：磷脂酰肌醇转移蛋白（PITP）在信号转导和膜运输中起作用[1，2]。旨在阐明PITP作用机制的研究产生了一个单一的主题。 PITP的活性源于其将磷脂酰肌醇（PI）从其合成位点转移到细胞活性位点以及刺激PI磷酸化形式的局部合成的能力。各种磷酸肌醇参与EGF信号转导和EGF受体的运输已得到充分证明[3]。使用荧光寿命成像显微镜（FLIM）来测量EGFP-PITP蛋白和荧光标记的磷脂之间的荧光共振能量转移（FRET），我们报道当用EGF刺激时，PITPalpha和PITPbeta可以与质膜上的PI或PC动态相互作用。另外，PITPbeta位于高尔基体，EGF刺激导致增强的FRET。 PLC的抑制剂和Ras / MAP激酶途径均能抑制EGF刺激的PITP（x与PI在质膜上的相互作用），通过光漂白后的荧光恢复（FRAP）测定PITP蛋白的迁移率，并EGF刺激降低了质膜的迁移率，我们得出的结论是PITPa。和PITPbeta在体内的动态行为是一个受调控的过程，涉及多种机制。

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