Institute for Integrated Cell-Material Sciences, Kyoto University, Nishikyo-ku, Kyoto, Japan.
Institute of Advanced Energy, Kyoto University, Uji, Kyoto, Japan.
Methods Mol Biol. 2020;2091:47-57. doi: 10.1007/978-1-0716-0167-9_4.
The pleckstrin homology (PH) domain is a family of structurally conserved proteins which can bind inositol phosphate derivatives. Some proteins involved in cellular signaling and cytoskeletal organization possess split PH domains that assemble into a structure which can bind specific inositol phosphates. Here we describe the design of split PH domain from a structurally well-characterized PH domain of phospholipase C (PLC) δ and Bruton's tyrosine kinase (Btk), which selectively bind Ins(1,4,5)P and Ins(1,3,4,5)P, respectively. The PH domains fold into a functional structure when the split halves are brought to close proximity, and can be utilized to detect specific inositol phosphate of interest.
pleckstrin 同源（PH）结构域是一类结构保守的蛋白家族，能够与肌醇磷酸衍生物结合。一些参与细胞信号转导和细胞骨架组织的蛋白拥有分裂 PH 结构域，这些分裂 PH 结构域组装成一个可以结合特定肌醇磷酸的结构。在此，我们描述了从结构上得到充分表征的 PLC δ 和 Bruton 酪氨酸激酶（Btk）的 PH 结构域设计，它们分别选择性地结合 Ins(1,4,5)P 和 Ins(1,3,4,5)P。当分裂的两半接近时，PH 结构域折叠成一个功能性结构，并可用于检测感兴趣的特定肌醇磷酸。
