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  • Title: Dealing with particles in different conformational states by electron microscopy and image processing.
    Author: Mellwig C, Böttcher B.
    Journal: J Struct Biol; 2001; 133(2-3):214-20. PubMed ID: 11472092.
    Abstract:
    Electron microscopy and image processing are powerful tools for investigating different conformational states of enzymes. It is not always possible to isolate these often unstable intermediates as single species. As a result electron micrographs show a snapshot of enzymes in various conformational states. We describe here how to recognize that the imaged particles have different conformations and how to obtain for each species a three-dimensional model using single-particle image processing. We investigated the ATP synthase from chloroplasts, which has a molecular mass of about 550 kDa. It is a membrane-bound enzyme and consists of two segments, a membrane-embedded hydrophobic F(0) part and a hydrophilic F(1) part. Analysis of the particle images indicated that the molecules were in two different conformations. For both conformations three-dimensional models were calculated, which showed that the structures differed mainly in the tilt of the F(0) part with respect to the F(1) part.
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