Imaging the Nanoscale Distribution of Phosphoinositides in the Cell Plasma Membrane with Single-Molecule Localization Super-Resolution Microscopy.
Imaging the Nanoscale Distribution of Phosphoinositides in the Cell Plasma Membrane with Single-Molecule Localization Super-Resolution Microscopy.
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DOI：
10.1007/978-1-0716-1142-5_6
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发表时间：
2021
影响因子：
--
通讯作者：
F. Fan;Chen Ji;X. Lou
中科院分区：
文献类型：
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作者：
F. Fan;Chen Ji;X. Lou
磷脂酰肌醇仅构成细胞磷脂的一小部分，但以基本方式控制细胞功能。通过蛋白质的相互作用，磷酸肌醇定义细胞器的身份和调节蛋白质的功能和组织和招聘在细胞溶质-膜界面。因此，对磷酸肌醇代谢的干扰改变细胞生理学并导致广泛的人类疾病，包括癌症和糖尿病。磷脂酰肌醇4，5-二磷酸（PtdIns（4，5）P2，又称PI（4，5）P2或PIP 2）是磷脂酰肌醇的主要成员。PtdIns（4，5）P2除了在磷脂酶C的第二信使途径中起作用外，还调节膜运输以及细胞骨架、离子通道和转运蛋白的功能。PtdIns（4，5）P2在质膜中的纳米级组织对于理解细胞信号在时间和空间上的特异性变得至关重要。在此，我们描述了一种单分子方法，通过使用超分辨率显微镜和基于PLCδ 1 pleckstrin同源（PH）结构域的双色荧光探针来可视化PtdIns（4，5）P2在质膜中的纳米级分布。这种方法可以扩展到图像其他磷酸肌醇通过改变特定的探针。
Phosphoinositides make up only a small fraction of cellular phospholipids yet control cell function in a fundamental manner. Through protein interactions, phosphoinositides define cellular organelle identity and regulate protein function and organization and recruitment at the cytosol–membrane interface. As a result, perturbations on phosphoinositide metabolism alter cell physiology and lead to a wide range of human diseases, including cancer and diabetes. Among seven phosphoinositide members, phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2, also known as PI(4,5)P2or PIP2) is abundant in the plasma membrane. Besides its role in the second messenger pathway of phospholipase C that cleaves PtdIns(4,5)P2to form diacylglycerol and inositol-1,4,5-trisphosphate (IP3), PtdIns(4,5)P2regulates membrane trafficking and the function of the cytoskeleton, ion channels, and transporters. The nanoscale organization of PtdIns(4,5)P2in the plasma membrane becomes essential to understand cellular signaling specificity in time and space. Here, we describe a single-molecule method to visualize the nanoscale distribution of PtdIns(4,5)P2in the plasma membrane by using super-resolution microscopy and the dual-color fluorescent probes based on the PLCδ1pleckstrin homology (PH) domain. This approach can be extended to image other phosphoinositides by changing the specific probes.