Abstract
Electron microscopy (EM) has made possible to solve the structure of many proteins. However, the resolution of some of the EM maps is too low for interpretation at the atomic level, which is particularly important to describe function. We describe methods that combine low-resolution EM data with atomic structures for different conformations of the same protein in order to produce atomic models compatible with the EM map.
We illustrate these methods with EM data from decavanadate-induced tubular crystals of a pseudo-phosphorylated intermediate of Ca-ATPase and the various atomic structures of other intermediates available in the Protein Data Bank (PDB). Determination of atomic structure permits not only to analyse protein–protein interactions in the crystals, but also to localize residues in the proximity of the crystallizing agent both within Ca-ATPase and between Ca-ATPase molecules.
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Acknowledgment
The authors would like to thank Professor D.L. Stokes for generously providing the EM map. This work was supported by CNRS (Centre National de la Recherche Scientifique) and CEA (Commissariat à l’Energie Atomique).
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Hinsen, K., Beaumont, E., Fournier, B., Lacapère, JJ. (2010). From Electron Microscopy Maps to Atomic Structures Using Normal Mode-Based Fitting. In: Lacapère, JJ. (eds) Membrane Protein Structure Determination. Methods in Molecular Biology, vol 654. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-762-4_13
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DOI: https://doi.org/10.1007/978-1-60761-762-4_13
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