Minimally disruptive optical control of protein tyrosine phosphatase 1B
Minimally disruptive optical control of protein tyrosine phosphatase 1B
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DOI：
10.1038/s41467-020-14567-8
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发表时间：
2020-02-07
影响因子：
16.6
通讯作者：
Fox, Jerome M.
中科院分区：
文献类型：
--
作者：
Hongdusit, Akarawin;Zwart, Peter H.;Fox, Jerome M.
蛋白酪氨酸磷酸酶调节大量重要的亚细胞信号事件，但它们仍然很难在其天然生物物理背景下进行研究。在这里，我们开发了一种最小干扰的光学方法来控制蛋白酪氨酸磷酸酶1B(PTP1B)--受体酪氨酸激酶的重要调节因子，也是治疗糖尿病、肥胖和癌症的靶标--我们使用这种方法来探索这种酶的细胞内功能。我们的保守的光控制体系结构，包括一个融合了变构调节元件的基于蛋白质的光开关，保留了PTP1B的天然结构、活性和亚细胞定位，提供了与细胞内翻译后修饰引起的活性变化相匹配的活性变化，并允许对光调制的分子基础进行实验分析。研究结果表明，最引人注目的是，PTP1B活性的微小变化可以导致其调控靶标的磷酸化状态发生重大变化。
Protein tyrosine phosphatases regulate a myriad of essential subcellular signaling events, yet they remain difficult to study in their native biophysical context. Here we develop a minimally disruptive optical approach to control protein tyrosine phosphatase 1B (PTP1B)-an important regulator of receptor tyrosine kinases and a therapeutic target for the treatment of diabetes, obesity, and cancer-and we use that approach to probe the intracellular function of this enzyme. Our conservative architecture for photocontrol, which consists of a protein-based light switch fused to an allosteric regulatory element, preserves the native structure, activity, and subcellular localization of PTP1B, affords changes in activity that match those elicited by post-translational modifications inside the cell, and permits experimental analyses of the molecular basis of optical modulation. Findings indicate, most strikingly, that small changes in the activity of PTP1B can cause large shifts in the phosphorylation states of its regulatory targets.