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Purification, crystallization and preliminary X‐ray diffraction analysis of RafE, a sugar‐binding lipoprotein from Streptococcus pneumoniae

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Streptococcus pneumoniae contains a large number of sugar‐transport systems and the system responsible for raffinose uptake has recently been identified. The substrate‐binding protein component of this system shares strong sequence homology with the multiple sugar metabolism substrate‐binding protein MsmE from S. mutans and contains a lipoprotein‐attachment site at cysteine residue 23. A truncated form (residues 24–419) of RafE from S. pneumoniae was cloned and overexpressed in Escherichia coli. Native and selenomethionine‐labelled protein have been crystallized in the hexagonal space group P6122. Diffraction data have been successfully phased to 2.90 Å using Se SAD data and model building is in progress.
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Keywords: RafE; SAD; SP1897; raffinose; ABC transporter; selenomethionine

Document Type: Research Article

Publication date: July 1, 2006

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