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Updated: Nov 3, 2025
Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
Published on: July 1, 2022
Gold nanoparticles and tilt pairs to assess protein flexibility by cryo-electron microscopy
Milind Jagota1, Raphael J L Townshend2, Lin-Woo Kang3
1Department of Computer Science, Stanford University, Stanford, CA, USA; Department of Electrical Engineering, Stanford University, Stanford, CA, USA.
Abstract:
A computational method was developed to recover the three-dimensional coordinates of gold nanoparticles specifically attached to a protein complex from tilt-pair images collected by electron microscopy. The program was tested on a simulated dataset and applied to a real dataset comprising tilt-pair images recorded by cryo electron microscopy of RNA polymerase II in a complex with four gold-labeled single-chain antibody fragments. The positions of the gold nanoparticles were determined, and comparison of the coordinates among the tetrameric particles revealed the range of motion within the protein complexes.
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