Jovanovic-Talisman Tijana, Chait Brian T, Rout Michael P
Department of Molecular Medicine, Beckman Research Institute of the City of Hope Comprehensive Cancer Center, Duarte, California, USA.
Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, USA.
Methods Cell Biol. 2014;122:379-93. doi: 10.1016/B978-0-12-417160-2.00017-5.
In vitro mimics of cellular machines have been recently engineered and utilized to investigate processes within cells. These devices can provide novel insights into biological mechanisms and have the potential to improve biotechnological processes such as separation. In particular, several devices have been designed to mimic translocation through nuclear pore complexes (NPCs). We describe here the fabrication of a biomimetic NPC using nanoporous filters lined with FG-repeats of proteins that create a selectivity barrier. We show the utility of this nanoselective filter as a testbed for the investigation of nucleocytoplasmic transport and demonstrate that this device closely reproduces key features of trafficking through the NPC.
近年来,人们设计并利用了细胞机器的体外模拟物来研究细胞内的过程。这些装置能够为生物学机制提供新的见解,并且具有改善诸如分离等生物技术过程的潜力。特别是,已经设计了几种装置来模拟通过核孔复合体(NPC)的转运。我们在此描述了一种仿生NPC的制造方法,该方法使用内衬蛋白质FG重复序列的纳米多孔过滤器来创建一个选择性屏障。我们展示了这种纳米选择性过滤器作为研究核质运输试验台的效用,并证明该装置紧密再现了通过NPC进行运输的关键特征。