Crystallization and preliminary X-ray analysis of the Streptomyces olivaceoviridis NgcE binding protein of the ABC transporter for N-acetylglucosamine.
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.
Saito
Akihiro
A
National Institute of Agrobiological Sciences, Universität Osnabrück, Osnabrück, Germany.
Fujimoto
Zui
Z
Minami
Eiichi
E
Mizuno
Hiroshi
H
Miyashita
Kiyotaka
K
Schrempf
Hildgund
H
Momma
Mitsuru
M
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
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D008024:Ligands; D012997:Solvents; D012645:Selenomethionine; D000117:Acetylglucosamine