Three-dimensional structure of proteins determined by electron microscopy

Recent developments in specimen preparation and image processing techniques have made it possible to determine the three-dimensional structure of proteins by electron microscopy. Periodic supramolecular aggregates of the protein under investigation are requiring to minimize radiation damage and to maximize the signal-to-noise ratio of structural detail. Useful information about the fine structure of the protein (e.g. binding sites for interacting molecules, antigenic determinants) can often be obtained by stoichiometric labeling of the ordered arrays with interacting molecules or antibody fragments, and computing difference maps from the reconstructions of the labeled and native structures. The use of this approach to molecular structure determination of proteins will be discussed in light of our work with bacteriophage and actin.