Particle fusion of super-resolution data reveals the unit structure of Nup96 in Nuclear Pore Complex

Particle fusion of super-resolution data reveals the unit structure of Nup96 in Nuclear Pore Complex

Wang, W. (TU Delft ImPhys/Computational Imaging)
Jakobi, A. (TU Delft BN/Arjen Jakobi Lab)
Wu, Yu-Le (European Molecular Biology Laboratory Heidelberg)
Ries, Jonas (University of Vienna)
Stallinga, S. (TU Delft ImPhys/Imaging Physics)
Rieger, B. (TU Delft ImPhys/Computational Imaging; TU Delft ImPhys/Rieger group)

ImPhys/Imaging Physics

2023

Single molecule localization microscopy offers resolution nearly down to the molecular level with specific molecular labelling, and is thereby a promising tool for structural biology. In practice, however, the actual value to this field is limited primarily by incomplete fluorescent labelling of the structure. This missing information can be completed by merging information from many structurally identical particles in a particle fusion approach similar to cryo-EM single-particle analysis. In this paper, we present a data analysis of particle fusion results of fluorescently labelled Nup96 nucleoporins in the Nuclear Pore Complex to show that Nup96 occurs in a spatial arrangement of two rings of 8 units with two Nup96 copies per unit giving a total of 32 Nup96 copies per pore. We use Artificial Intelligence assisted modeling in Alphafold to extend the existing cryo-EM model of Nup96 to accurately pinpoint the positions of the fluorescent labels and show the accuracy of the match between fluorescent and cryo-EM data to be better than 3 nm in-plane and 5 nm out-of-plane.

http://resolver.tudelft.nl/uuid:1b72b2c7-c148-48be-b019-c03935d3f7f1

https://doi.org/10.1038/s41598-023-39829-5

2045-2322

Scientific Reports, 13 (1)

Institutional Repository

journal article

© 2023 W. Wang, A. Jakobi, Yu-Le Wu, Jonas Ries, S. Stallinga, B. Rieger