Open AccessStructural analysis of non‐crystalline macromolecules: the ribosome
Author(s) -
Orlova Elena V.
Publication year - 2000
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s090744490001115x
Subject(s) - macromolecule , electron microscope , single particle analysis , cryo electron microscopy , resolution (logic) , microscopy , ribosome , particle (ecology) , biological system , materials science , chemistry , nanotechnology , crystallography , physics , optics , computer science , biology , artificial intelligence , biochemistry , organic chemistry , aerosol , ecology , rna , gene
Single‐particle analysis using cryo‐electron microscopy has emerged recently as a tool for elucidating the structure of biological macromolecules and their assemblies. A prerequisite for single‐particle analysis is an ensemble of images of structurally identical particles in different orientations. There are a variety of techniques used for image processing of this type of object in electron microscopy. The paper gives an overview of the general philosophy of image analysis of single particles in electron microscopy. It has been shown that multivariate statistical analysis of large data sets in conjunction with angular reconstitution is capable of yielding a structural resolution approaching that of X‐ray structural analysis of large macromolecules. Structure preservation during specimen preparation for electron microscopy is crucial for high‐resolution studies. Nowadays, cryo‐electron microscopy is a beneficial method for studying biological macromolecules in their natural environment, allowing their rapid freezing in a particular functional state. The processing of images of 50S Escherichia coli ribosomal subunits embedded in vitreous ice is used as an example of image analysis of single particles at 7.5 Å resolution.