Cloning, purification and crystallization of Bacillus anthracis class C acid phosphatase
Cloning, expression, purification and crystallization studies of a recombinant class C acid phosphatase from the Category A pathogen Bacillus anthracis are reported. Large diffraction‐quality crystals were grown in the presence of HEPES and Jeffamine ED‐2001 at pH 7.0. The crystals belong to space group P212121, with unit‐cell parameters a = 53.4, b = 90.1, c = 104.2 Å. The asymmetric unit is predicted to contain two protein molecules with a solvent content of 38%. Two native data sets were collected from the same crystal before and after flash‐annealing. The first data set had a mosaicity of 1.6° and a high‐resolution limit of 1.8 Å. After flash‐annealing, the apparent mosaicity decreased to 0.9° and the high‐resolution limit of usable data increased to 1.6 Å. This crystal form is currently being used to determine the structure of B. anthracis class C acid phosphatase with experimental phasing techniques.
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