Investigating molecular crowding within nuclear pores using polarization-PALM. Academic Article uri icon


  • The key component of the nuclear pore complex (NPC) controlling permeability, selectivity, and the speed of nucleocytoplasmic transport is an assembly of natively unfolded polypeptides, which contain phenylalanine-glycine (FG) binding sites for nuclear transport receptors. The architecture and dynamics of the FG-network have been refractory to characterization due to the paucity of experimental methods able to probe the mobility and density of the FG-polypeptides and embedded macromolecules within intact NPCs. Combining fluorescence polarization, super-resolution microscopy, and mathematical analyses, we examined the rotational mobility of fluorescent probes at various locations within the FG-network under different conditions. We demonstrate that polarization PALM (p-PALM) provides a rich source of information about low rotational mobilities that are inaccessible with bulk fluorescence anisotropy approaches, and anticipate that p-PALM is well-suited to explore numerous crowded cellular environments. In total, our findings indicate that the NPC's internal organization consists of multiple dynamic environments with different local properties.

published proceedings

  • Elife

altmetric score

  • 1

author list (cited authors)

  • Fu, G., Tu, L., Zilman, A., & Musser, S. M.

citation count

  • 12

complete list of authors

  • Fu, Guo||Tu, Li-Chun||Zilman, Anton||Musser, Siegfried M

publication date

  • September 2017


published in