Utility of Butvar support film and methylamine tungstate stain in three-dimensional electron microscopy: agreement between stain and frozen-hydrated reconstructions

Journal of Structural Biology
S J KolodziejJ K Stoops

Abstract

A random conical tilt reconstruction of negatively stained Saccharomyces cerevisiae fatty acid synthase was used as a model to compute a three-dimensional reconstruction from untilted stain specimens of the molecules in multiple orientations using a three-dimensional projection alignment method. The resulting structure (24 A resolution) has a more uniform resolution than the initial structure and the handedness revealed in the random conical tilt method is preserved. In a similar approach, this model was used to compute a 21-A-resolution frozen-hydrated structure from untilted specimens of the molecules in multiple orientations. Even though the reconstructions are in close agreement, the stain structure appears to enhance the protein density associated with less robust features. These procedures significantly reduce the time and effort required to obtain a three-dimensional reconstruction from frozen-hydrated data with a resolution that is comparable to the best obtained by more laborious methods. The agreement between the stain and frozen-hydrated reconstructions affords convincing evidence concerning the validity of the structure and the information afforded by the two reconstructions significantly enhances the structural ana...Continue Reading

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Citations

Dec 9, 1998·Journal of Structural Biology·S J KolodziejJ K Stoops
Dec 16, 2000·Trends in Biochemical Sciences·W Baumeister, A C Steven
Apr 27, 2002·Micron : the International Research and Review Journal for Microscopy·J Robin Harris, Dirk Scheffler
Jul 31, 1998·Micron : the International Research and Review Journal for Microscopy·M AdrianJ R Harris
Aug 25, 2010·Quarterly Reviews of Biophysics·Timm MaierNenad Ban
Mar 1, 2012·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Andrea FeraThomas S Reese
Dec 21, 2007·Acta Crystallographica. Section D, Biological Crystallography·Yong Xiong
Jan 28, 2009·Acta Crystallographica. Section D, Biological Crystallography·Simon Jenni, Nenad Ban
Jun 23, 2021·Chemical Reviews·Pedro PaivaPedro A Fernandes
Apr 25, 2006·Chemistry & Biology·Craig A TownsendTsion Bililign
Mar 4, 2006·Science·Simon JenniNenad Ban

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