Schifferer Martina, Feng Suihan, Stein Frank, Tischer Christian, Schultz Carsten
European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
Bioorg Med Chem. 2015 Jun 15;23(12):2862-7. doi: 10.1016/j.bmc.2015.03.048. Epub 2015 Mar 25.
Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP2 sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP2 recovery. This permits the observation of the PIP2-dependent clathrin assembly at the PM.
化学二聚体是用于在完整细胞内使分子聚集的强大非侵入性工具。我们最近引入了一种快速可逆的化学二聚体系统，该系统能够使酶在质膜（PM）与细胞质之间进行瞬时转运。在此，我们应用该系统通过磷酸肌醇5-磷酸酶（5Ptase）的转运在质膜处瞬时激活磷脂酰肌醇4,5-二磷酸（PIP2）的分解。我们发现，加入可逆化学二聚体rCD1后，PIP2传感器磷脂酶C-δ PH结构域（PLCδ-PH）从质膜释放。通过与rCD1竞争，5Ptase从质膜快速释放后PIP2得以恢复。这使得能够观察质膜处依赖PIP2的网格蛋白组装。
