Crystallization and preliminary X-ray analysis of FliJ, a cytoplasmic component of the flagellar type III protein-export apparatus from Salmonella sp.
The axial component proteins of the bacterial flagellum are synthesized in the cytoplasm and then translocated into the central channel of the flagellum by the flagellar type III protein-export apparatus for self-assembly at the distal growing end of the flagellum. FliJ is an essential cytoplasmic component of the export apparatus. In this study, Salmonella FliJ with an extra three residues (glycine, serine and histidine) attached to the N-terminus as the remainder of a His tag (GSH-FliJ) was purified and crystallized. Crystals were obtained by the sitting-drop vapour-diffusion technique using PEG 300 as a precipitant. GSH-FliJ crystals grew in the hexagonal space group P6(1)22 or P6(5)22. While the native crystals diffracted to 3.3 A resolution, the diffraction resolution limit of mercury derivatives was extended to 2.1 A. Anomalous and isomorphous difference Patterson maps of the mercury-derivative crystal showed significant peaks in their Harker sections, indicating the usefulness of the derivative data for structure determination.
Ibuki
Tatsuya
T
Graduate School of Frontier Biosciences, Osaka University, Suita, Japan.
Shimada
Masafumi
M
Minamino
Tohru
T
Namba
Keiichi
K
Imada
Katsumi
K
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D018360:Crystallography, X-Ray / Q000379:methods*; D003593:Cytoplasm / Q000378:metabolism; D005407:Flagella / Q000378:metabolism*; D005978:Glutathione / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D008628:Mercury / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012475:Salmonella / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; C069253:fliJ protein, Bacteria; D012694:Serine; D006639:Histidine; D008628:Mercury; D005978:Glutathione; D005998:Glycine