Other Structure Determination Methods
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There are more ways of gaining insight into macromolecular structure than X-ray diffraction. Like X-ray diffraction, some of these are based on the generation of ordered arrays of the molecule to be studied. For many reasons, based on either the protein or its function, this is not always possible. Others, some of which are currently enjoying a marked increase in popularity, do not require crystals. Many of these come with the added advantage that they can be used to capture reaction intermediates and/or enable the experimenter to observe changes in specific amino acids, which is often not possible with X-ray diffraction methods. This chapter divides into two sections; those methods that can be used to obtain a 3D structure (neutron diffraction, cryogenic electron microscopy, nuclear magnetic resonance, and X-ray free electron laser diffraction) and those that are suitable for more general structural information (chemical cross linking, fluorescence resonance energy transfer, circular dichroism). Virtually all of the methods discussed below can be expanded for the study of other aspects of macromolecular structure-function relationships, and some, such as fluorescence and chemical cross linking, are a subset of a rich methodology for the study of macromolecules.
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