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Title
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Selenomethionine labeling of large biological macromolecular complexes: probing the structure of marine bacterial virus PM2.
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Authors
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H.M.Kivelä,
N.G.Abrescia,
J.K.Bamford,
J.M.Grimes,
D.I.Stuart,
D.H.Bamford.
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Ref.
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J Struct Biol, 2008,
161,
204-210.
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PubMed id
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Abstract
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There is a need for improved tools for labeling protein species within large
macromolecular assemblies. Here we describe a method for the efficient
selenomethionine labeling of the membrane-containing bacterial virus PM2 for
structural studies. By examining potential host cells a strain was found which
was auxotrophic for methionine, and by performing a multiparameter search of
conditions it was possible to derive a robust protocol which simultaneously
minimized the toxic effects of the selenomethionine, so that a reasonable virus
yield was maintained, whilst still achieving essentially complete labeling. This
has allowed us to fingerprint the protein constituents of the virus in a
relatively low resolution electron density map. Such a technique can be adapted
to other macromolecule complexes studied by X-ray crystallography.
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