Cloning, purification, crystallization and preliminary crystallographic studies of Bradyrhizobium fucosyltransferase NodZ

Authors: Brzezinski, Krzysztof; Rogozinski, Bartosz; Stepkowski, Tomasz; Bujacz, Grzegorz; Jaskolski, Mariusz

Source: Acta Crystallographica Section D, Volume 60, Number 2, 1 February 2004 , pp. 344-346(3)

Publisher: Blackwell Publishing

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Abstract:

The α-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 P6₁22 or P6₅22, with unit-cell parameters a = 125.5, c = 95.6 Å, and contain 56.8% solvent and a single protein molecule in the asymmetric unit. Native data were collected to 2.85 Å using synchrotron radiation and cryogenic conditions. The native crystals were soaked in a mother-liquor solution containing 2.5 mM [Ta₆Br₁₂]²⁺ cluster for derivatization and SAD data were collected to 3.4 Å at the tantalum L_III absorption peak.

Keywords: fucosyltransferase; Nod factor; nodulation; nitogen fixation; symbiosis; [Ta6Br12]2+ cluster

Document Type: Research article

DOI: 10.1107/S0907444903027227

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