title: Crystallization and preliminary X-ray analysis of the Streptomyces olivaceoviridis NgcE binding protein of the ABC transporter for N-acetylglucosamine.
abstract: The NgcE protein binds N-acetylglucosamine (GIcNAc) as well asN,N'-diacetylchitobiose and is a component of the ABC transporter Ngc for GIcNAc uptake in Streptomyces olivaceoviridis. After cloning the corresponding gene in an Escherichia coli host, the NgcE protein was overproduced in a soluble form within the cytoplasm and purified to homogeneity by four consecutive chromatographic processes. Crystals of NgcE that grew in the presence of 1 mM GlcNAc,20%(w/v) PEG MME 2000 and 100 mM Tris-HCI pH 8.5 had a plate-like shape and belonged to either space group P21212 (unit-cell parameters a = 59.9, b = 153.0, c = 41.7 A) or P212121 (a = 58.1, b = 96.3, c = 151.7 A). The former crystals diffracted to 1.8 A resolution andthe latter to 2.2 A. Selenomethionine-containing crystals were generated under the same conditions and belonged to space group P212121 with unit-cell parameters a = 58.4, b = 96.6, c = 152.5 A, and diffracted to 2.0 A resolution.
authors.affiliation: National Institute of Agrobiological Sciences, Universität Osnabrück, Osnabrück, Germany.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Saito
authors.forename: Zui
authors.initials: Z
authors.lastname: Fujimoto
authors.forename: Eiichi
authors.initials: E
authors.lastname: Minami
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mizuno
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Miyashita
authors.forename: Hildgund
authors.initials: H
authors.lastname: Schrempf
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Momma
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry*; D000117:Acetylglucosamine / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D002845:Chromatography; D003001:Cloning, Molecular; D003461:Crystallography; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D008024:Ligands; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012645:Selenomethionine / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013302:Streptomyces / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D008024:Ligands; D012997:Solvents; D012645:Selenomethionine; D000117:Acetylglucosamine