Zernike Phase Plate Cryoelectron Microscopy Facilitates Single Particle Analysis of Unstained Asymmetric Protein Complexes

Wei Hau Chang, Michael T K Chiu, Chin Yu Chen, Chi Fu Yen, Yen Cheng Lin, Yi Ping Weng, Ji Chau Chang, Yi Min Wu, Holland Cheng, Jianhua Fu, I. Ping Tu

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Single particle reconstruction from cryoelectron microscopy images, though emerging as a powerful means in structural biology, is faced with challenges as applied to asymmetric proteins smaller than megadaltons due to low contrast. Zernike phase plate can improve the contrast by restoring the microscope contrast transfer function. Here, by exploiting simulated Zernike and conventional defocused cryoelectron microscope images with noise characteristics comparable to those of experimental data, we quantified the efficiencies of the steps in single particle analysis of ice-embedded RNA polymerase II (500 kDa), transferrin receptor complex (290 kDa), and T7 RNA polymerase lysozyme (100 kDa). Our results show Zernike phase plate imaging is more effective as to particle identification and also sorting of orientations, conformations, and compositions. Moreover, our analysis on image alignment indicates that Zernike phase plate can, in principle, reduce the number of particles required to attain near atomic resolution by 10-100 fold for proteins between 100 kDa and 500 kDa.

Original languageEnglish (US)
Pages (from-to)17-27
Number of pages11
JournalStructure
Volume18
Issue number1
DOIs
StatePublished - Jan 13 2010

Keywords

  • PROTEINS

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Fingerprint Dive into the research topics of 'Zernike Phase Plate Cryoelectron Microscopy Facilitates Single Particle Analysis of Unstained Asymmetric Protein Complexes'. Together they form a unique fingerprint.

  • Cite this

    Chang, W. H., Chiu, M. T. K., Chen, C. Y., Yen, C. F., Lin, Y. C., Weng, Y. P., Chang, J. C., Wu, Y. M., Cheng, H., Fu, J., & Tu, I. P. (2010). Zernike Phase Plate Cryoelectron Microscopy Facilitates Single Particle Analysis of Unstained Asymmetric Protein Complexes. Structure, 18(1), 17-27. https://doi.org/10.1016/j.str.2009.12.001