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PMID: 10096920 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Solution x-ray scattering-based estimation of electron cryomicroscopy imaging parameters for reconstruction of virus particles.

Biophysical journal ·Vol. 76 ·No. 4 ·1999-04-00 ·Pages 2249-61

Thuman-Commike PA, Tsuruta H, Greene B, Prevelige PE, King J, Chiu W

Abstract

Structure factor amplitudes and phases can be computed directly from electron cryomicroscopy images. Inherent aberrations of the electromagnetic lenses and other instrumental factors affect the structure factors, however, resulting in decreased accuracy in the determined three-dimensional reconstruction. In contrast, solution x-ray scattering provides absolute and accurate measurement of spherically averaged structure factor amplitudes of particles in solution but does not provide information on the phases. In the present study, we explore the merits of using solution x-ray scattering data to estimate the imaging parameters necessary to make corrections to the structure factor amplitudes derived from electron cryomicroscopic images of icosahedral virus particles. Using 400-kV spot-scan images of the bacteriophage P22 procapsid, we have calculated an amplitude contrast of 8.0 +/- 5.2%. The amplitude decay parameter has been estimated to be 523 +/- 188 A2 with image noise compensation and 44 +/- 66 A2 without it. These results can also be used to estimate the minimum number of virus particles needed for reconstruction at different resolutions.

MeSH Terms
Bacteriophage P22/ultrastructure Biophysical Phenomena Biophysics Capsid/ultrastructure Cryoelectron Microscopy/methods,statistics & numerical data Image Processing, Computer-Assisted/methods,statistics & numerical data Scattering, Radiation Solutions
Chemicals
Solutions
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Thuman-Commike P A
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030, USA.
Tsuruta H
Greene B
Prevelige P E
King J
Chiu W
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(99)77381-9
Published
1999-04-00
Pages
2249-61
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1300198
Subset
IM
Grants
NIGMS NIH HHS · GM17980 · United States
NIGMS NIH HHS · GM47980 · United States
NCRR NIH HHS · P41RR02250 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(99)77381-9 PMC Full Text View journal details
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