Spatial filtering of electron micrographs of negatively stained alpha-amylase crystals

Images of the alpha-amylase molecule from pig pancreas have been obtained in two projections by the application of spatial filtering techniques to electron micrographs of negatively stained microcrystals of the enzyme. The Fourier filtering was performed on a PPD 11/40 digital computer after microdensitometry on an Optronics P-1000 photoscan system. The set of procedures and programs we employed are described herein. We compared these images with equivalent images obtained by three-dimensional X-ray diffraction analysis using conventional isomorphous replacement at approximately twice the resolution. We find that there is quite good agreement between the two kinds of images, and that a number of gross structural features compare quite well. We conclude that the combination of electron microscopy with digital spatial filtering when applied to protein microcrystals yields very respectable results.