Nuclear Pore Scaffold Structure

Very interesting. Fluorescence microscopy combined with particle averaging to attain a precision below one nanometer; used to look at the organization of a macromolecular structure: the nuclear pore.

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Published Online July 11 2013Science 9 August 2013: 
Vol. 341 no. 6146 pp. 655-658 
DOI: 10.1126/science.1240672
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Nuclear Pore Scaffold Structure Analyzed by Super-Resolution Microscopy and Particle Averaging

1. Anna Szymborska¹, 
2. Alex de Marco², 
3. Nathalie Daigle¹, 
4. Volker C. Cordes³, 
5. John A. G. Briggs²,
6. Jan Ellenberg¹,*

+Author Affiliations

1. ¹Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
2. ²Structural and Computational Biology Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
3. ³Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.

1. ↵*Corresponding author. E-mail: jan.ellenberg@embl.de

• ABSTRACT
• EDITOR'S SUMMARY

Much of life’s essential molecular machinery consists of large protein assemblies that currently pose challenges for structure determination. A prominent example is the nuclear pore complex (NPC), for which the organization of its individual components remains unknown. By combining stochastic super-resolution microscopy, to directly resolve the ringlike structure of the NPC, with single particle averaging, to use information from thousands of pores, we determined the average positions of fluorescent molecular labels in the NPC with a precision well below 1 nanometer. Applying this approach systematically to the largest building block of the NPC, the Nup107-160 subcomplex, we assessed the structure of the NPC scaffold. Thus, light microscopy can be used to study the molecular organization of large protein complexes in situ in whole cells.

Science Article