Cloning, purification, crystallization and preliminary crystallographic studies of Bradyrhizobium fucosyltransferase NodZ.
The alpha-1,6-fucosyltransferase NodZ from Bradyrhizobium sp. WM9 (Lupinus), composed of 325 amino acids with a molecular weight of 37 kDa, has been cloned, expressed and purified. Protein crystals suitable for X-ray diffraction were obtained under optimized crystallization conditions using ammonium dihydrogen phosphate as a precipitant. The crystals are hexagonal and belong to space group P6(1)22 or P6(5)22, with unit-cell parameters a = 125.5, c = 95.6 A, and contain 56.8% solvent and a single protein molecule in the asymmetric unit. Native data were collected to 2.85 A using synchrotron radiation and cryogenic conditions. The native crystals were soaked in a mother-liquor solution containing 2.5 mM [Ta(6)Br(12)](2+) cluster for derivatization and SAD data were collected to 3.4 A at the tantalum L(III) absorption peak.
Brzezinski
Krzysztof
K
Department of Crystallography, Faculty of Chemistry, A. Mickiewicz University, Grunwaldzka 6, 60-780 Poznan, Poland.
Rogozinski
Bartosz
B
Stepkowski
Tomasz
T
Bujacz
Grzegorz
G
Jaskolski
Mariusz
M
D001426:Bacterial Proteins / Q000737:chemistry*; D020369:Bradyrhizobium / Q000201:enzymology*; D003001:Cloning, Molecular; D005647:Fucosyltransferases / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D017356:Synchrotrons; D013696:Temperature; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D010710:Phosphates; D000644:Quaternary Ammonium Compounds; D005647:Fucosyltransferases; C103941:NodZ protein, bacteria