title: Purification, crystallization and preliminary X-ray diffraction analysis of RafE, a sugar-binding lipoprotein from Streptococcus pneumoniae.
abstract: 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 P6(1)22. Diffraction data have been successfully phased to 2.90 angstroms using Se SAD data and model building is in progress.
authors.affiliation: Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA, Scotland. neison@chem.gla.ac.uk
authors.forename: Neil G
authors.initials: NG
authors.lastname: Paterson
authors.forename: Alan
authors.initials: A
authors.lastname: Riboldi-Tunnicliffe
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Mitchell
authors.forename: Neil W
authors.initials: NW
authors.lastname: Isaacs
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D003461:Crystallography; D004926:Escherichia coli / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D011887:Raffinose / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012645:Selenomethionine / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D008074:Lipoproteins; D011994:Recombinant Proteins; D012645:Selenomethionine; D011887:Raffinose