Flemming Dirk, Stelter Philipp, Hurt Ed
Biochemie-Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
Methods Cell Biol. 2014;122:99-115. doi: 10.1016/B978-0-12-417160-2.00005-9.
The discovery of dynein light chain 2 (Dyn2) as a member of the nucleoporins in yeast led to a series of applications to study NPC structure and function. Its intriguing ability to act as a hub for the parallel dimerization of two short amino acid sequence motifs (DID) prompted us to utilize it as a tool for probing nucleocytoplasmic transport in vivo. Further, the distinct structure of the Dyn2-DID rod, which is easily visible in the electron microscope, allowed us to develop a precise structural label on proteins or protein complexes. This label was used to identify the position of subunits in NPC subcomplexes or to derive at pseudo-atomic models of single large Nups. The versatility for various applications of the DID-Dyn2 system makes it an attractive molecular tool beyond the nuclear pore and transport field.
动力蛋白轻链2（Dyn2）作为酵母核孔蛋白成员的发现引发了一系列用于研究核孔复合体（NPC）结构和功能的应用。它具有作为两个短氨基酸序列基序（DID）平行二聚化中心的有趣能力，这促使我们将其用作探测体内核质运输的工具。此外，Dyn2-DID杆的独特结构在电子显微镜下很容易看到，这使我们能够在蛋白质或蛋白质复合物上开发精确的结构标记。该标记用于确定NPC亚复合物中亚基的位置或推导单个大核孔蛋白的伪原子模型。DID-Dyn2系统在各种应用中的多功能性使其成为核孔和运输领域之外有吸引力的分子工具。
